PRMT5 inhibitors and uses thereof

ABSTRACT

Described herein are compounds of Formula (A), pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof. Compounds of the present invention are useful for inhibiting PRMT5 activity. Methods of using the compounds for treating PRMT5-mediated disorders are also described.

RELATED APPLICATIONS

The present application is a national stage filing under 35 U.S.C. §371of International PCT application PCT/US2013/077221, filed Dec. 20, 2013which claims priority under 35 U.S.C. §119(e) to U.S. provisional patentapplications, U.S. Ser. No. 61/745,477, filed Dec. 21, 2012, and U.S.Ser. No. 61/785,025, filed Mar. 14, 2013, the entire contents of each ofwhich are incorporated herein by reference.

BACKGROUND OF THE INVENTION

Epigenetic regulation of gene expression is an important biologicaldeterminant of protein production and cellular differentiation and playsa significant pathogenic role in a number of human diseases.

Epigenetic regulation involves heritable modification of geneticmaterial without changing its nucleotide sequence. Typically, epigeneticregulation is mediated by selective and reversible modification (e.g.,methylation) of DNA and proteins (e.g., histones) that control theconformational transition between transcriptionally active and inactivestates of chromatin. These covalent modifications can be controlled byenzymes such as methyltransferases (e.g., PRMT5), many of which areassociated with specific genetic alterations that can cause humandisease.

Disease-associated chromatin-modifying enzymes (e.g., PRMT5) play a rolein diseases such as proliferative disorders, metabolic disorders, andblood disorders. Thus, there is a need for the development of smallmolecules that are capable of inhibiting the activity of PRMT5.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Protein arginine methyltransferase 5 (PRMT5) catalyzes the addition oftwo methyl groups to the two w-guanidino nitrogen atoms of arginine,resulting in ω-NG, N′G symmetric dimethylation of arginine (sDMA) of thetarget protein. PRMT5 functions in the nucleus as well as in thecytoplasm, and its substrates include histones, spliceosomal proteins,transcription factors (See e.g., Sun et al., PNAS (2011) 108:20538-20543). PRMT5 generally functions as part of a molecule weightprotein complex. While the protein complexes of PRMT5 can have a varietyof components, they generally include the protein MEP50 (methylosomeprotein 50). In addition, PRMT5 acts in conjunction with cofactor SAM(S-adenosyl methionine).

PRMT5 is an attractive target for modulation given its role in theregulation of diverse biological processes. It has now been found thatcompounds described herein, and pharmaceutically acceptable salts andcompositions thereof, are effective as inhibitors of PRMT5.

Such compounds have the general Formula (A):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R, R⁵,R⁶, R⁷, R⁸, R^(x), R¹², R¹³, and n are as defined herein.

In some embodiments, the inhibitors of PRMT5 have the general Formula(I):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R, R⁵,R⁶, R⁷, R⁸, R^(x), and n are as defined herein.

In some embodiments, pharmaceutical compositions are provided whichcomprise a compound described herein (e.g., a compound of Formula (A),e.g., Formula (I)), or a pharmaceutically acceptable salt thereof, andoptionally a pharmaceutically acceptable excipient.

In certain embodiments, compounds described herein inhibit activity ofPRMT5. In certain embodiments, methods of inhibiting PRMT5 are providedwhich comprise contacting PRMT5 with an effective amount of a compoundof Formula (A), e.g., Formula (I), or a pharmaceutically acceptable saltthereof. The PRMT5 may be purified or crude, and may be present in acell, tissue, or a subject. Thus, such methods encompass inhibition ofPRMT5 activity both in vitro and in vivo. In certain embodiments, thePRMT5 is wild-type PRMT5. In certain embodiments, the PRMT5 isoverexpressed. In certain embodiments, the PRMT5 is a mutant. In certainembodiments, the PRMT5 is in a cell. In certain embodiments, the PRMT5is in an animal, e.g., a human. In some embodiments, the PRMT5 is in asubject that is susceptible to normal levels of PRMT5 activity due toone or more mutations associated with a PRMT5 substrate. In someembodiments, the PRMT5 is in a subject known or identified as havingabnormal PRMT5 activity (e.g., overexpression). In some embodiments, aprovided compound is selective for PRMT5 over other methyltransferases.In certain embodiments, a provided compound is at least about 10-foldselective, at least about 20-fold selective, at least about 30-foldselective, at least about 40-fold selective, at least about 50-foldselective, at least about 60-fold selective, at least about 70-foldselective, at least about 80-fold selective, at least about 90-foldselective, or at least about 100-fold selective relative to one or moreother methyltransferases.

In certain embodiments, methods of altering gene expression in a cellare provided which comprise contacting a cell with an effective amountof a compound of Formula (A), e.g., Formula (I), or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition thereof. Incertain embodiments, the cell in culture in vitro. In certainembodiments, cell is in an animal, e.g., a human.

In certain embodiments, methods of altering transcription in a cell areprovided which comprise contacting a cell with an effective amount of acompound of Formula (A), e.g., Formula (I), or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition thereof. Incertain embodiments, the cell in culture in vitro. In certainembodiments, the cell is in an animal, e.g., a human.

In some embodiments, methods of treating a PRMT5-mediated disorder areprovided which comprise administering to a subject suffering from aPRMT5-mediated disorder an effective amount of a compound describedherein (e.g., a compound of Formula (A), e.g., Formula (I)), or apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition thereof. In certain embodiments, the PRMT5-mediated disorderis a proliferative disorder, a metabolic disorder, or a blood disorder.In certain embodiments, compounds described herein are useful fortreating cancer. In certain embodiments, compounds described herein areuseful for treating hematopoietic cancer, lung cancer, prostate cancer,melanoma, or pancreatic cancer. In certain embodiments, compoundsdescribed herein are useful for treating a hemoglobinopathy. In certainembodiments, compounds described herein are useful for treating sicklecell anemia. In certain embodiments, compounds described herein areuseful for treating diabetes or obesity. In certain embodiments, aprovided compound is useful in treating inflammatory and autoimmunedisease.

Compounds described herein are also useful for the study of PRMT5 inbiological and pathological phenomena, the study of intracellular signaltransduction pathways mediated by PRMT5, and the comparative evaluationof new PRMT5 inhibitors.

This application refers to various issued patent, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition,John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, 1989; and Carruthers,Some Modern Methods of Organic Synthesis, 3^(rd) Edition, CambridgeUniversity Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers,and thus can exist in various isomeric forms, e.g., enantiomers and/ordiastereomers. For example, the compounds described herein can be in theform of an individual enantiomer, diastereomer or geometric isomer, orcan be in the form of a mixture of stereoisomers, including racemicmixtures and mixtures enriched in one or more stereoisomer. Isomers canbe isolated from mixtures by methods known to those skilled in the art,including chiral high pressure liquid chromatography (HPLC) and theformation and crystallization of chiral salts; or preferred isomers canbe prepared by asymmetric syntheses. See, for example, Jacques et al.,Enantiomers, Racemates and Resolutions (Wiley Interscience, New York,1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistryof Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables ofResolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ.of Notre Dame Press, Notre Dame, Ind. 1972). The present disclosureadditionally encompasses compounds described herein as individualisomers substantially free of other isomers, and alternatively, asmixtures of various isomers.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofany compound described herein does not exclude any tautomer form.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds that differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of hydrogen by deuterium ortritium, replacement of ¹⁹F with ¹⁸F, or the replacement of a carbon bya ¹³C- or ¹⁴C-enriched carbon are within the scope of the disclosure.Such compounds are useful, for example, as analytical tools or probes inbiological assays.

The term “aliphatic,” as used herein, includes both saturated andunsaturated, nonaromatic, straight chain (i.e., unbranched), branched,acyclic, and cyclic (i.e., carbocyclic) hydrocarbons. In someembodiments, an aliphatic group is optionally substituted with one ormore functional groups. As will be appreciated by one of ordinary skillin the art, “aliphatic” is intended herein to include alkyl, alkenyl,alkynyl, cycloalkyl, and cycloalkenyl moieties.

When a range of values is listed, it is intended to encompass each valueand sub-range within the range. For example “C₁₋₆ alkyl” is intended toencompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆,C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄, C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

“Alkyl” refers to a radical of a straight-chain or branched saturatedhydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀ alkyl”). Insome embodiments, an alkyl group has 1 to 10 carbon atoms (“C₁₋₁₀alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms(“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1 to 8 carbonatoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl group has 1 to 7carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkyl group has 1to 6 carbon atoms (“C₁₋₆ alkyl”). In some embodiments, an alkyl grouphas 1 to 5 carbon atoms (“C₁₋₅ alkyl”). In some embodiments, an alkylgroup has 1 to 4 carbon atoms (“C₁₋₄ alkyl”). In some embodiments, analkyl group has 1 to 3 carbon atoms (“C₁₋₃ alkyl”). In some embodiments,an alkyl group has 1 to 2 carbon atoms (“C₁₋₂ alkyl”). In someembodiments, an alkyl group has 1 carbon atom (“C₁ alkyl”). In someembodiments, an alkyl group has 2 to 6 carbon atoms (“C₂₋₆ alkyl”).Examples of C₁₋₆ alkyl groups include methyl (C₁), ethyl (C₂), n-propyl(C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄),iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl(C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈)and the like. In certain embodiments, each instance of an alkyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents. In certain embodiments, the alkyl group isunsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments, thealkyl group is substituted C₁₋₁₀ alkyl.

In some embodiments, an alkyl group is substituted with one or morehalogens. “Perhaloalkyl” is a substituted alkyl group as defined hereinwherein all of the hydrogen atoms are independently replaced by ahalogen, e.g., fluoro, bromo, chloro, or iodo. In some embodiments, thealkyl moiety has 1 to 8 carbon atoms (“C₁₋₈ perhaloalkyl”). In someembodiments, the alkyl moiety has 1 to 6 carbon atoms (“C₁₋₆perhaloalkyl”). In some embodiments, the alkyl moiety has 1 to 4 carbonatoms (“C₁₋₄ perhaloalkyl”). In some embodiments, the alkyl moiety has 1to 3 carbon atoms (“C₁₋₃ perhaloalkyl”). In some embodiments, the alkylmoiety has 1 to 2 carbon atoms (“C₁₋₂ perhaloalkyl”). In someembodiments, all of the hydrogen atoms are replaced with fluoro. In someembodiments, all of the hydrogen atoms are replaced with chloro.Examples of perhaloalkyl groups include —CF₃, —CF₂CF₃, —CF₂CF₂CF₃,—CCl₃, —CFCl₂, —CF₂Cl, and the like.

“Alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon double bonds, and no triple bonds (“C₂₋₂₀ alkenyl”). Insome embodiments, an alkenyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms(“C₂₋₉ alkenyl”). In some embodiments, an alkenyl group has 2 to 8carbon atoms (“C₂₋₈ alkenyl”). In some embodiments, an alkenyl group has2 to 7 carbon atoms (“C₂₋₇ alkenyl”). In some embodiments, an alkenylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkenyl”). In some embodiments, analkenyl group has 2 to 5 carbon atoms (“C₂₋₅ alkenyl”). In someembodiments, an alkenyl group has 2 to 4 carbon atoms (“C₂₋₄ alkenyl”).In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C₂₋₃alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C₂alkenyl”). The one or more carbon-carbon double bonds can be internal(such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples ofC₂₋₄ alkenyl groups include ethenyl (C₂), 1-propenyl (C₃), 2-propenyl(C₃), 1-butenyl (C₄), 2-butenyl (C₄), butadienyl (C₄), and the like.Examples of C₂₋₆ alkenyl groups include the aforementioned C₂₋₄ alkenylgroups as well as pentenyl (C₅), pentadienyl (C₅), hexenyl (C₆), and thelike. Additional examples of alkenyl include heptenyl (C₇), octenyl(C₈), octatrienyl (C₈), and the like. In certain embodiments, eachinstance of an alkenyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkenyl”) or substituted (a“substituted alkenyl”) with one or more substituents. In certainembodiments, the alkenyl group is unsubstituted C₂₋₁₀ alkenyl. Incertain embodiments, the alkenyl group is substituted C₂₋₁₀ alkenyl.

“Alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon triple bonds, and optionally one or more double bonds(“C₂₋₂₀ alkynyl”). In some embodiments, an alkynyl group has 2 to 10carbon atoms (“C₂₋₁₀ alkynyl”). In some embodiments, an alkynyl grouphas 2 to 9 carbon atoms (“C₂₋₉ alkynyl”). In some embodiments, analkynyl group has 2 to 8 carbon atoms (“C₂₋₈ alkynyl”). In someembodiments, an alkynyl group has 2 to 7 carbon atoms (“C₂-7 alkynyl”).In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C₂₋₆alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms(“C₂₋₅ alkynyl”). In some embodiments, an alkynyl group has 2 to 4carbon atoms (“C₂₋₄ alkynyl”). In some embodiments, an alkynyl group has2 to 3 carbon atoms (“C₂₋₃ alkynyl”). In some embodiments, an alkynylgroup has 2 carbon atoms (“C₂ alkynyl”). The one or more carbon-carbontriple bonds can be internal (such as in 2-butynyl) or terminal (such asin 1-butynyl). Examples of C₂₋₄ alkynyl groups include, withoutlimitation, ethynyl (C₂), 1-propynyl (C₃), 2-propynyl (C₃), 1-butynyl(C₄), 2-butynyl (C₄), and the like. Examples of C₂-alkenyl groupsinclude the aforementioned C₂₋₄ alkynyl groups as well as pentynyl (C₅),hexynyl (C₆), and the like. Additional examples of alkynyl includeheptynyl (C₇), octynyl (C₈), and the like. In certain embodiments, eachinstance of an alkynyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents. In certainembodiments, the alkynyl group is unsubstituted C₂₋₁₀ alkynyl. Incertain embodiments, the alkynyl group is substituted C₂₋₁₀ alkynyl.

“Carbocyclyl” or “carbocyclic” refers to a radical of a non-aromaticcyclic hydrocarbon group having from 3 to 14 ring carbon atoms (“C₃₋₁₄carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. Insome embodiments, a carbocyclyl group has 3 to 10 ring carbon atoms(“C₃₋₁₀ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to8 ring carbon atoms (“C₃₋₈ carbocyclyl”). In some embodiments, acarbocyclyl group has 3 to 6 ring carbon atoms (“C₃-6 carbocyclyl”). Insome embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms(“C₃-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to10 ring carbon atoms (“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclylgroups include, without limitation, cyclopropyl (C₃), cyclopropenyl(C₃), cyclobutyl (C₄), cyclobutenyl (C₄), cyclopentyl (C₅),cyclopentenyl (C₅), cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl(C₆), and the like. Exemplary C₃₋₈ carbocyclyl groups include, withoutlimitation, the aforementioned C₃₋₆ carbocyclyl groups as well ascycloheptyl (C₇), cycloheptenyl (C₇), cycloheptadienyl (C₇),cycloheptatrienyl (C₇), cyclooctyl (C₈), cyclooctenyl (C₈),bicyclo[2.2.1]heptanyl (C₇), bicyclo[2.2.2]octanyl (C₈), and the like.Exemplary C₃₋₁₀ carbocyclyl groups include, without limitation, theaforementioned C₃₋₈ carbocyclyl groups as well as cyclononyl (C₉),cyclononenyl (C₉), cyclodecyl (C₁₀), cyclodecenyl (C₁₀),octahydro-1H-indenyl (C₉), decahydronaphthalenyl (C₁₀),spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examplesillustrate, in certain embodiments, the carbocyclyl group is eithermonocyclic (“monocyclic carbocyclyl”) or is a fused, bridged or spiroring system such as a bicyclic system (“bicyclic carbocyclyl”) and canbe saturated or can be partially unsaturated. “Carbocyclyl” alsoincludes ring systems wherein the carbocyclyl ring, as defined above, isfused with one or more aryl or heteroaryl groups wherein the point ofattachment is on the carbocyclyl ring, and in such instances, the numberof carbons continue to designate the number of carbons in thecarbocyclic ring system. In certain embodiments, each instance of acarbocyclyl group is independently optionally substituted, e.g.,unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a“substituted carbocyclyl”) with one or more substituents. In certainembodiments, the carbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl group is a substituted C₃₋₁₀carbocyclyl.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 14 ring carbon atoms (“C₃₋₁₄cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 10 ringcarbon atoms (“C₃₋₁₀ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 8 ring carbon atoms (“C₃₋₈ cycloalkyl”). In someembodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C₃₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ringcarbon atoms (“C₅₋₆ cycloalkyl”). In some embodiments, a cycloalkylgroup has 5 to 10 ring carbon atoms (“C₅₋₁₀ cycloalkyl”). Examples ofC₅₋₆ cycloalkyl groups include cyclopentyl (C₅) and cyclohexyl (C₅).Examples of C₃₋₆ cycloalkyl groups include the aforementioned C₅₋₆cycloalkyl groups as well as cyclopropyl (C₃) and cyclobutyl (C₄).Examples of C₃ 8 cycloalkyl groups include the aforementioned C₃₋₆cycloalkyl groups as well as cycloheptyl (C₇) and cyclooctyl (C₈). Incertain embodiments, each instance of a cycloalkyl group isindependently unsubstituted (an “unsubstituted cycloalkyl”) orsubstituted (a “substituted cycloalkyl”) with one or more substituents.In certain embodiments, the cycloalkyl group is unsubstituted C₃₋₁₀cycloalkyl. In certain embodiments, the cycloalkyl group is substitutedC₃₋₁₀ cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3- to14-membered non-aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-14 membered heterocyclyl”). In certainembodiments, heterocyclyl or heterocyclic refers to a radical of a 3-10membered non-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). Inheterocyclyl groups that contain one or more nitrogen atoms, the pointof attachment can be a carbon or nitrogen atom, as valency permits. Aheterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”)or a fused, bridged or spiro ring system such as a bicyclic system(“bicyclic heterocyclyl”), and can be saturated or can be partiallyunsaturated. Heterocyclyl bicyclic ring systems can include one or moreheteroatoms in one or both rings. “Heterocyclyl” also includes ringsystems wherein the heterocyclyl ring, as defined above, is fused withone or more carbocyclyl groups wherein the point of attachment is eitheron the carbocyclyl or heterocyclyl ring, or ring systems wherein theheterocyclyl ring, as defined above, is fused with one or more aryl orheteroaryl groups, wherein the point of attachment is on theheterocyclyl ring, and in such instances, the number of ring memberscontinue to designate the number of ring members in the heterocyclylring system. In certain embodiments, each instance of heterocyclyl isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substitutedheterocyclyl”) with one or more substituents. In certain embodiments,the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. Incertain embodiments, the heterocyclyl group is substituted 3-10 memberedheterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 memberednon-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-10 membered heterocyclyl”). In someembodiments, a heterocyclyl group is a 5-8 membered non-aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms, wherein eachheteroatom is independently selected from nitrogen, oxygen, and sulfur(“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl groupis a 5-6 membered non-aromatic ring system having ring carbon atoms and1-4 ring heteroatoms, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In someembodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has one ring heteroatomselected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude, without limitation, azirdinyl, oxiranyl, and thiorenyl.Exemplary 4-membered heterocyclyl groups containing one heteroatominclude, without limitation, azetidinyl, oxetanyl, and thietanyl.Exemplary 5-membered heterocyclyl groups containing one heteroatominclude, without limitation, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl,and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, dioxolanyl,oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-memberedheterocyclyl groups containing three heteroatoms include, withoutlimitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary6-membered heterocyclyl groups containing one heteroatom include,without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include, without limitation, piperazinyl, morpholinyl,dithianyl, and dioxanyl. Exemplary 6-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, triazinanyl.Exemplary 7-membered heterocyclyl groups containing one heteroatominclude, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom include,without limitation, azocanyl, oxecanyl, and thiocanyl. Exemplary5-membered heterocyclyl groups fused to a C₆ aryl ring (also referred toherein as a 5,6-bicyclic heterocyclic ring) include, without limitation,indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,benzoxazolinonyl, and the like.

Exemplary 6-membered heterocyclyl groups fused to an aryl ring (alsoreferred to herein as a 6,6-bicyclic heterocyclic ring) include, withoutlimitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclicor tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 itelectrons shared in a cyclic array) having 6-14 ring carbon atoms andzero heteroatoms provided in the aromatic ring system (“C₆₋₁₄ aryl”). Insome embodiments, an aryl group has six ring carbon atoms (“C₆ aryl”;e.g., phenyl). In some embodiments, an aryl group has ten ring carbonatoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). Insome embodiments, an aryl group has fourteen ring carbon atoms (“C₁₄aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein thearyl ring, as defined above, is fused with one or more carbocyclyl orheterocyclyl groups wherein the radical or point of attachment is on thearyl ring, and in such instances, the number of carbon atoms continue todesignate the number of carbon atoms in the aryl ring system. In certainembodiments, each instance of an aryl group is independently optionallysubstituted, e.g., unsubstituted (an “unsubstituted aryl”) orsubstituted (a “substituted aryl”) with one or more substituents. Incertain embodiments, the aryl group is unsubstituted C₆₋₁₄ aryl. Incertain embodiments, the aryl group is substituted C₆₋₁₄ aryl.

“Heteroaryl” refers to a radical of a 5-14 membered monocyclic orpolycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system(e.g., having 6 or 10 it electrons shared in a cyclic array) having ringcarbon atoms and 1-4 ring heteroatoms provided in the aromatic ringsystem, wherein each heteroatom is independently selected from nitrogen,oxygen and sulfur (“5-14 membered heteroaryl”). In certain embodiments,heteroaryl refers to a radical of a 5-10 membered monocyclic or bicyclic4n+2 aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, e.g., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-14 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-14 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-10 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, each instanceof a heteroaryl group is independently optionally substituted, e.g.,unsubstituted (“unsubstituted heteroaryl”) or substituted (“substitutedheteroaryl”) with one or more substituents. In certain embodiments, theheteroaryl group is unsubstituted 5-14 membered heteroaryl. In certainembodiments, the heteroaryl group is substituted 5-14 memberedheteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatominclude, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary5-membered heteroaryl groups containing two heteroatoms include, withoutlimitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, andisothiazolyl. Exemplary 5-membered heteroaryl groups containing threeheteroatoms include, without limitation, triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include, without limitation, tetrazolyl. Exemplary6-membered heteroaryl groups containing one heteroatom include, withoutlimitation, pyridinyl. Exemplary 6-membered heteroaryl groups containingtwo heteroatoms include, without limitation, pyridazinyl, pyrimidinyl,and pyrazinyl. Exemplary 6-membered heteroaryl groups containing threeor four heteroatoms include, without limitation, triazinyl andtetrazinyl, respectively. Exemplary 7-membered heteroaryl groupscontaining one heteroatom include, without limitation, azepinyl,oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groupsinclude, without limitation, indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude, without limitation, naphthyridinyl, pteridinyl, quinolinyl,isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

“Fused” or “ortho-fused” are used interchangeably herein, and refer totwo rings that have two atoms and one bond in common, e.g.,

“Bridged” refers to a ring system containing (1) a bridgehead atom orgroup of atoms which connect two or more non-adjacent positions of thesame ring; or (2) a bridgehead atom or group of atoms which connect twoor more positions of different rings of a ring system and does notthereby form an ortho-fused ring, e.g.,

“Spiro” or “Spiro-fused” refers to a group of atoms which connect to thesame atom of a carbocyclic or heterocyclic ring system (geminalattachment), thereby forming a ring, e.g.,

Spiro-fusion at a bridgehead atom is also contemplated.

“Partially unsaturated” refers to a group that includes at least onedouble or triple bond. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aromatic groups (e.g., aryl or heteroaryl groups) asherein defined. Likewise, “saturated” refers to a group that does notcontain a double or triple bond, i.e., contains all single bonds.

In some embodiments, aliphatic, alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl groups, as defined herein, areoptionally substituted (e.g., “substituted” or “unsubstituted”aliphatic, “substituted” or “unsubstituted” alkyl, “substituted” or“unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl,“substituted” or “unsubstituted” carbocyclyl, “substituted” or“unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or“substituted” or “unsubstituted” heteroaryl group). In general, the term“substituted”, whether preceded by the term “optionally” or not, meansthat at least one hydrogen present on a group (e.g., a carbon ornitrogen atom) is replaced with a permissible substituent, e.g., asubstituent which upon substitution results in a stable compound, e.g.,a compound which does not spontaneously undergo transformation such asby rearrangement, cyclization, elimination, or other reaction. Unlessotherwise indicated, a “substituted” group has a substituent at one ormore substitutable positions of the group, and when more than oneposition in any given structure is substituted, the substituent iseither the same or different at each position. The term “substituted” iscontemplated to include substitution with all permissible substituentsof organic compounds, including any of the substituents described hereinthat results in the formation of a stable compound. The presentdisclosure contemplates any and all such combinations in order to arriveat a stable compound. For purposes of this disclosure, heteroatoms suchas nitrogen may have hydrogen substituents and/or any suitablesubstituent as described herein which satisfy the valencies of theheteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to,halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂,—N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻, —N(OR^(cc))R^(bb), —SH, —SR^(aa),—SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO, —C(OR^(cc))₂, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —OC(═NR^(bb))R^(aa),—OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —OC(═NR^(bb))N(R^(bb))₂,—NR^(bb)C(═NR^(bb))N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), —SO₂OR^(aa), —OSO₂R^(aa),—S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃,—OSi(R^(aa))₃—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa),—SC(═S)SR^(aa), —SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa),—SC(═O)R^(aa), —P(═O)₂R^(aa), —OP(═O)₂R^(aa), —P(═O)(R^(aa))₂,—OP(═O)(R^(aa))₂, —OP(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂,—OP(═O)₂N(R^(bb))₂, —P(═O)(NR^(bb))₂, —OP(═O)(NR^(bb))₂,—NR^(bb)P(═O)(OR^(cc))₂, —NR^(bb)P(═O)(NR^(bb))₂, —P(R^(cc))₂,—P(R^(cc))₃, —OP(R^(cc))₂, —OP(R^(cc))₃, —B(R^(aa))₂, —B(OR^(cc))₂,—BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

or two geminal hydrogens on a carbon atom are replaced with the group═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R^(aa), ═NNR^(bb)C(═O)OR^(aa),═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or ═NOR^(cc);

each instance of R^(aa) is, independently, selected from C₁₋₁₀ alkyl,C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, ortwo R^(aa) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(bb) is, independently, selected from hydrogen, —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R, —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂,—SO₂R^(cc), —SO₂OR, —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc),—C(═S)SR^(cc), —P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂,—P(═O)(NR^(cc))₂, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(bb) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(cc) is, independently, selected from hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups are joined to form a 3-14 memberedheterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

-   -   each instance of R^(dd) is, independently, selected from        halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee),        —ON(R^(ff))₂, —N(R^(ff))₂, —N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff),        —SH, —SR^(ee), —SSR^(ee), —C(═O)R^(ee), —CO₂H, —CO₂R^(ee),        —OC(═O)R^(ee), —OCO₂R^(ee), —C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂,        —NR^(ff)C(═O)R^(ee), —NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂,        —C(═NR^(ff))OR^(ee), OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee),        —C(═NR^(ff))N(R^(ff))₂, —OC(═NR^(ff))N(R^(ff))₂,        —NR^(ff)C(═NR^(ff))N(R^(ff))₂, —NR^(ff)SO₂R^(ee),        —SO₂N(R^(ff))₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),        —S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,        —C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)₂R^(ee),        —P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₆        alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀        carbocyclyl, 3-10 membered heterocyclyl, C₆₋₁₀ aryl, 5-10        membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,        carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently        substituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups, or two        geminal R^(dd) substituents can be joined to form ═O or ═S;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups;

each instance of R^(ff) is, independently, selected from hydrogen, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl,3-10 membered heterocyclyl, C₆₋₁₀ aryl and 5-10 membered heteroaryl, ortwo R^(ff) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; and

each instance of R^(gg) is, independently, halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆ alkyl)₂,—N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)⁺X⁻, —NH₃⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁₋₆ alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃-C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)₂(C₁₋₆alkyl), —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₁₀ carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R^(gg) substituents can be joined to form ═Oor ═S; wherein X is a counterion.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a cationic quaternary amino group in order to maintainelectronic neutrality. Exemplary counterions include halide ions (e.g.,F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, sulfonate ions(e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate,benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate,naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonicacid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate,ethanoate, propanoate, benzoate, glycerate, lactate, tartrate,glycolate, and the like).

“Halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro,—Cl), bromine (bromo, —Br), or iodine (iodo, —I).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quarternary nitrogenatoms. Exemplary nitrogen atom substitutents include, but are notlimited to, hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(bb))R^(aa),—C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc),—SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂, —P(═O)(NR^(cc))₂,C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups attached to a nitrogen atom are joinedto form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc) and R^(dd) are asdefined above.

In certain embodiments, the substituent present on a nitrogen atom is anitrogen protecting group (also referred to as an amino protectinggroup). Nitrogen protecting groups include, but are not limited to, —OH,—OR^(aa), —N(R^(cc))₂, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))R^(aa), —C(═NR^(cc))OR^(aa),—C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc),—SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc), C₁₋₁₀ alkyl(e.g., aralkyl, heteroaralkyl), C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aralkyl, aryl, and heteroaryl is independently substitutedwith 0, 1, 2, 3, 4, or 5 R^(dd) groups, and wherein R^(aa), R^(bb),R^(cc), and R^(dd) are as defined herein. Nitrogen protecting groups arewell known in the art and include those described in detail inProtecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts,3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Amide nitrogen protecting groups (e.g., —C(═O)R^(aa)) include, but arenot limited to, formamide, acetamide, chloroacetamide,trichloroacetamide, trifluoroacetamide, phenylacetamide,3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide,N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide,o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,(N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide,3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine,o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Carbamate nitrogen protecting groups (e.g., —C(═O)OR^(aa)) include, butare not limited to, methyl carbamate, ethyl carbamante,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzylcarbamate.

Sulfonamide nitrogen protecting groups (e.g., —S(═O)₂R^(aa)) include,but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to,phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacylderivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanylderivative, N-acetylmethionine derivative,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate,N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is anoxygen protecting group (also referred to as a hydroxyl protectinggroup). Oxygen protecting groups include, but are not limited to,—R^(aa), —N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa),—C(═NR^(bb))N(R^(bb))₂, —S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃,—P(R^(cc))₂, —P(R^(cc))₃, —P(═O)₂R^(aa), —P(═O)(R^(aa))₂,—P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and —P(═O)(NR^(bb))₂, whereinR^(aa), R^(bb), and R^(cc) are as defined herein. Oxygen protectinggroups are well known in the art and include those described in detailin Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.Wuts, 3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Exemplary oxygen protecting groups include, but are not limited to,methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxyl)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), t-butyl carbonate (BOC), alkylmethyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethylcarbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc),2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethylcarbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkylisobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts).

In certain embodiments, the substituent present on a sulfur atom is asulfur protecting group (also referred to as a thiol protecting group).Sulfur protecting groups include, but are not limited to, —R^(aa),—N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃,—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and—P(═O)(NR^(bb))₂, wherein R^(aa), R^(bb), and R^(cc) are as definedherein. Sulfur protecting groups are well known in the art and includethose described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999,incorporated herein by reference.

As used herein, a “leaving group”, or “LG”, is a term understood in theart to referred to a molecular fragment that departs with a pair ofelectrons upon heterolytic bond cleavage, wherein the molecular fragmentis an anion or neutral molecule. See, for example, Smith, March AdvancedOrganic Chemistry 6th ed. (501-502). Examples of suitable leaving groupsinclude, but are not limited to, halides (such as chloride, bromide, oriodide), alkoxycarbonyloxy, aryloxycarbonyloxy, alkanesulfonyloxy,arenesulfonyloxy, alkyl-carbonyloxy (e.g., acetoxy), arylcarbonyloxy,aryloxy, methoxy, N,O-dimethylhydroxylamino, pixyl, haloformates, —NO₂,trialkylammonium, and aryliodonium salts. In some embodiments, theleaving group is a sulfonic acid ester. In some embodiments, thesulfonic acid ester comprises the formula —OSO₂R^(LG1) wherein R^(LG1)is selected from the group consisting alkyl optionally, alkenyloptionally substituted, heteroalkyl optionally substituted, aryloptionally substituted, heteroaryl optionally substituted, arylalkyloptionally substituted, and heterarylalkyl optionally substituted. Insome embodiments, R^(LG1) is substituted or unsubstituted C₁-C₆ alkyl.In some embodiments, R is methyl. In some embodiments, R^(LG1) is —CF₃.In some embodiments, R^(LG1) is substituted or unsubstituted aryl. Insome embodiments, R^(LG1) is substituted or unsubstituted phenyl. Insome embodiments R^(LG1) is:

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, and claims. The present disclosureis not intended to be limited in any manner by the above exemplarylisting of substituents.

“Pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and other animals without undue toxicity,irritation, allergic response, and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, Berge et al. describepharmaceutically acceptable salts in detail in J. PharmaceuticalSciences (1977) 66:1-19. Pharmaceutically acceptable salts of thecompounds describe herein include those derived from suitable inorganicand organic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid, or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representativealkali or alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, quaternary salts.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (e.g., a male or female of any age group, e.g., apediatric subject (e.g, infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or othernon-human animals, for example, non-human mammals (e.g., primates (e.g.,cynomolgus monkeys, rhesus monkeys); commercially relevant mammals suchas cattle, pigs, horses, sheep, goats, cats, and/or dogs), birds (e.g.,commercially relevant birds such as chickens, ducks, geese, and/orturkeys), rodents (e.g., rats and/or mice), reptiles, amphibians, andfish. In certain embodiments, the non-human animal is a mammal. Thenon-human animal may be a male or female at any stage of development. Anon-human animal may be a transgenic animal.

“Condition,” “disease,” and “disorder” are used interchangeably herein.

“Treat,” “treating” and “treatment” encompasses an action that occurswhile a subject is suffering from a condition which reduces the severityof the condition or retards or slows the progression of the condition(“therapeutic treatment”). “Treat,” “treating” and “treatment” alsoencompasses an action that occurs before a subject begins to suffer fromthe condition and which inhibits or reduces the severity of thecondition (“prophylactic treatment”).

An “effective amount” of a compound refers to an amount sufficient toelicit the desired biological response, e.g., treat the condition. Aswill be appreciated by those of ordinary skill in this art, theeffective amount of a compound described herein may vary depending onsuch factors as the desired biological endpoint, the pharmacokinetics ofthe compound, the condition being treated, the mode of administration,and the age and health of the subject. An effective amount encompassestherapeutic and prophylactic treatment.

A “therapeutically effective amount” of a compound is an amountsufficient to provide a therapeutic benefit in the treatment of acondition or to delay or minimize one or more symptoms associated withthe condition. A therapeutically effective amount of a compound means anamount of therapeutic agent, alone or in combination with othertherapies, which provides a therapeutic benefit in the treatment of thecondition. The term “therapeutically effective amount” can encompass anamount that improves overall therapy, reduces or avoids symptoms orcauses of the condition, or enhances the therapeutic efficacy of anothertherapeutic agent.

A “prophylactically effective amount” of a compound is an amountsufficient to prevent a condition, or one or more symptoms associatedwith the condition or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of a therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the condition. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

As used herein, the term “methyltransferase” represents transferaseclass enzymes that are able to transfer a methyl group from a donormolecule to an acceptor molecule, e.g., an amino acid residue of aprotein or a nucleic base of a DNA molecule. Methytransferases typicallyuse a reactive methyl group bound to sulfur in S-adenosyl methionine(SAM) as the methyl donor. In some embodiments, a methyltransferasedescribed herein is a protein methyltransferase. In some embodiments, amethyltransferase described herein is a histone methyltransferase.Histone methyltransferases (HMT) are histone-modifying enzymes,(including histone-lysine N-methyltransferase and histone-arginineN-methyltransferase), that catalyze the transfer of one or more methylgroups to lysine and arginine residues of histone proteins. In certainembodiments, a methyltransferase described herein is a histone-arginineN-methyltransferase.

As generally described above, provided herein are compounds useful asPRMT5 inhibitors. In some embodiments, the present disclosure provides acompound of Formula (A):

or a pharmaceutically acceptable salt thereof,wherein

represents a single or double bond;

R¹² is hydrogen, halogen, or optionally substituted C₁₋₃alkyl;

R¹³ is hydrogen, halogen, optionally substituted C₁₋₃alkyl,—NR^(A1)R^(A2), or —OR¹;

R^(A1) and R^(A2) are each independently hydrogen, optionallysubstituted C₁₋₃ alkyl, a nitrogen protecting group, or R^(A1) andR^(A2) are taken together with the intervening nitrogen atom to form anoptionally substituted 3-6 membered heterocyclic ring;

Ring A is an optionally substituted, 5- to 12-membered, monocyclic orbicyclic, heterocyclyl or heteroaryl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur;

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

Y is O or S;

R⁵, R⁶, R⁷, and R⁸ are each independently hydrogen, halo, or optionallysubstituted aliphatic;

-   -   each R^(x) is independently selected from the group consisting        of halo, —CN, optionally substituted aliphatic, —OR′, and        —N(R^(aa))₂;

R′ is hydrogen or optionally substituted aliphatic;

-   -   each R″ is independently hydrogen or optionally substituted        aliphatic, or two R″ are taken together with their intervening        atoms to form a heterocyclic ring; and

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits.

In some embodiments, the carbon attached to R¹² has (S)-stereochemistry.In some embodiments, the carbon attached to R¹² has (R)-stereochemistry.In some embodiments, the carbon attached to R¹³ has (S)-stereochemistry.In some embodiments, the carbon attached to R¹³ has (R)-stereochemistry.In some embodiments, R¹² is hydrogen. In some embodiments, R¹³ ishydrogen. In some embodiments, both R¹² and R¹³ are hydrogen. In someembodiments, R¹² is optionally substituted C₁₋₃ alkyl. In someembodiments, R¹³ is optionally substituted C₁₋₃ alkyl. In someembodiments, both R¹² and R¹³ are optionally substituted C₁₋₃alkyl. Insome embodiments, R is halogen e.g., fluoro, bromo, chloro, or iodo,provided that R is not —OR¹. In some embodiments, R is halogen e.g.,fluoro, bromo, chloro, or iodo. In some embodiments, both R¹² and R¹³are halogen e.g., fluoro, bromo, chloro, or iodo. In some embodiments,R¹² is halogen e.g., fluoro, bromo, chloro, or iodo and R¹³ isoptionally substituted C₁₋₃alkyl. In some embodiments, R¹² is optionallysubstituted C₁₋₃alkyl and R¹³ is halogen e.g., fluoro, bromo, chloro, oriodo. In some embodiments, R¹³ is —OR¹. In some embodiments, R¹² isoptionally substituted C₁₋₃alkyl and R¹³ is —OR¹. In some embodiments,R¹² is hydrogen and R¹³ is —OR¹. In some embodiments, R¹² is hydrogenand R¹³ optionally substituted C₁₋₃alkyl. In some embodiments, R¹² isoptionally substituted C₁₋₃alkyl and R is hydrogen. In some embodiments,R is halogen e.g., fluoro, bromo, chloro, or iodo and R¹³ is hydrogen.In some embodiments, R¹² is hydrogen and R¹³ is halogen e.g., fluoro,bromo, chloro, or iodo.

As generally defined above, R¹² is hydrogen, halogen, or optionallysubstituted C₁₋₃alkyl. In certain embodiments, R¹² is hydrogen. Incertain embodiments, R¹² is optionally substituted C₁₋₃alkyl, e.g.,optionally substituted with halogen. In certain embodiments, R¹² isoptionally substituted C₁ alkyl, e.g., methyl or trifluoromethyl. Incertain embodiments, R¹² is optionally substituted C₂ alkyl, e.g.,ethyl. In certain embodiments, R is optionally substituted C₃ alkyl,e.g., propyl. In certain embodiments, R¹² is fluoro, provided that R isnot —OR¹. In certain embodiments, R is chloro, provided that R³ is not—OR¹. In certain embodiments, R¹² is bromo, provided that R is not —OR¹.In certain embodiments, R¹² is iodo, provided that R¹³ is not —OR¹.

As generally defined above, R¹³ is hydrogen, halogen, optionallysubstituted C₁₋₃alkyl, —NR^(A1)R^(A2) or —OR¹. In certain embodiments,R¹³ is hydrogen. In certain embodiments, R¹³ is optionally substitutedC₁₋₃alkyl, e.g., optionally substituted with halogen. In certainembodiments, R¹³ is optionally substituted C₁alkyl, e.g., methyl ortrifluoromethyl. In certain embodiments, R¹³ is optionally substitutedC₂ alkyl, e.g., ethyl. In certain embodiments, R is optionallysubstituted C₃ alkyl, e.g., propyl. In certain embodiments, R¹³ isfluoro. In certain embodiments, R is chloro. In certain embodiments, Ris bromo. In certain embodiments, R¹³ is iodo.

For example, in some embodiments of Formula (A), wherein R¹³ ishydrogen, the present disclosure provides a compound of Formula (A-1):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R⁵,R⁶, R⁷, R⁸, R^(x), R¹², and n are as described herein.

In some embodiments of Formula (A), wherein R¹² is hydrogen, the presentdisclosure provides a compound of Formula (A-2):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R⁵,R⁶, R⁷, R⁸, R^(x), R¹³, and n are as described herein.

In some embodiments of Formula (A), wherein both R¹² and R¹³ arehydrogen, the present disclosure provides a compound of Formula (A-3):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R⁵,R⁶, R⁷, R⁸, R^(x), and n are as described herein.

In some embodiments of Formula (A), wherein R¹³ is —OR¹, the presentdisclosure provides a compound of Formula (A-4):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R, R⁵,R⁶, R⁷, R⁸, R^(x), R¹², and n are as described herein.

In some embodiments of Formula (A), wherein R¹³ is —NR^(A1)R^(A2), thepresent disclosure provides a compound of Formula (A-5):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R⁵,R⁶, R⁷, R, R^(x), R², R^(A1), and R^(A2), and n are as described herein.

In some embodiments, the present disclosure provides a compound ofFormula (I):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R, R⁵,R⁶, R⁷, R⁸, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (I-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R¹,R⁵, R⁶, R⁷, R⁸, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (I-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R, R⁵,R⁶, R⁷, R⁸, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (I-c):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R¹,R^(x), and n are as described herein.

In some embodiments of Formula (A), the present disclosure provides acompound of Formula (A-6):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R^(x),R¹², R¹³, and n are as described herein.

In certain embodiments, a provided compound is of Formula (I′):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R¹,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (I′-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R¹,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (I′-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, Y, R¹,R^(x), and n are as described herein.

In some embodiments of Formula (A), the present disclosure provides acompound of Formula (A-7):

or a pharmaceutically acceptable salt thereof, wherein Y, R^(x), and nare as described herein,

G is NR², CR³R⁴, O or S;

R¹² is hydrogen, halogen, or optionally substituted C₁₋₃alkyl;

R¹³ is hydrogen, halogen, optionally substituted C₁₋₃alkyl,—NR^(A1)R^(A2), or —OR¹;

R^(A1) and R^(A2) are independently hydrogen, optionally substitutedC₁₋₃ alkyl, a nitrogen protecting group, or R^(A1) and R^(A2) are takentogether with the intervening nitrogen atom to form an optionallysubstituted 3-6 membered heterocyclic ring;

R¹ is hydrogen, R^(z), or —C(O)R^(z), wherein R^(z) is optionallysubstituted C₁₋₆ alkyl;

R² is selected from the group consisting of optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —C(O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A), —C(═S)N(R^(B))₂,—S(═O)R^(A), —SO₂R^(A), and —SO₂N(R^(B))₂;

R³ is selected from the group consisting of hydrogen, halo, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, and optionally substituted heteroaryl, or two R^(B) groups aretaken together with their intervening atoms to form an optionallysubstituted heterocyclic ring;

R⁴ is selected from the group consisting of hydrogen, halo, andoptionally substituted aliphatic;

each R^(y) is independently selected from the group consisting of halo,—CN, —NO₂, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, or two adjacent R^(y) groups may betaken together with their intervening atoms to form a saturated,partially unsaturated, or aromatic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur;

p is 0, 1, or 2; and

m is 0, 1, 2, 3, or 4.

In certain embodiments, a provided compound is of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein R¹, G, Y, R^(y),m, p, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (II-a):

or a pharmaceutically acceptable salt thereof, wherein R¹, G, Y, R^(y),m, p, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (II-b):

or a pharmaceutically acceptable salt thereof, wherein R¹, G, Y, R^(y),m, p, R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (A-8):

or a pharmaceutically acceptable salt thereof, wherein R², R^(y), m,R^(x), R¹², R¹³, and n are as described herein.

In certain embodiments, a provided compound is of Formula (III):

or a pharmaceutically acceptable salt thereof, wherein R², R^(y), m,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (III-a):

or a pharmaceutically acceptable salt thereof, wherein R², R^(y), m,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (III-b):

or a pharmaceutically acceptable salt thereof, wherein R², R^(y), m,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (A-9):

or a pharmaceutically acceptable salt thereof, wherein R³, R^(y), m,R^(x), R¹², R¹³, and n are as described herein.

In certain embodiments, a provided compound is of Formula (IV):

or a pharmaceutically acceptable salt thereof, wherein R³, R^(y), m,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (IV-a):

or a pharmaceutically acceptable salt thereof, wherein R³, R^(y), m,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (IV-b):

or a pharmaceutically acceptable salt thereof, wherein R³, R^(y), m,R^(x), and n are as described herein.

In certain embodiments, a provided compound is of Formula (A-10):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),R¹², R¹³, and n are as described herein.

In certain embodiments, a provided compound is of Formula (V):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (V-a):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (V-b):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (A-11):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),R¹², R¹³ and n are as described herein.

In certain embodiments, a provided compound is of Formula (VI):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (VI-a):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),and n are as described herein.

In certain embodiments, a provided compound is of Formula (VI-b):

or a pharmaceutically acceptable salt thereof, wherein R^(y), m, R^(x),and n are as described herein.

In some embodiments,

represents a single bond. In some embodiments,

represents a double bond.

As defined generally above, Ring A is an optionally substituted, 5- to12-membered, monocyclic or bicyclic, heterocyclyl or heteroaryl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. One of ordinary skill in the art will understand that Ring Acomprises an amide or thioamide. In certain embodiments, Ring A is anoptionally substituted, 5- to 6-membered, monocyclic heteroaryl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Ring A is an optionally substituted, 5-to 7-membered, monocyclic heterocyclyl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In certainembodiments, Ring A is an optionally substituted, 8- to 10-membered,bicyclic heteroaryl having 1-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, Ring A is anoptionally substituted, 8- to 12-membered, bicyclic heterocyclyl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, Ring A is an optionally substitutedpiperdinone. In certain embodiments, Ring A is an optionally substitutedpyridinone. In certain embodiments, Ring A is an optionally substitutedpiperazinone. In certain embodiments, Ring A is an optionallysubstituted isoindolinone. In certain embodiments, Ring A is anoptionally substituted 2H-benzo[b][1,4]oxazin-3(4H)-one. In someembodiments, Ring A is

wherein G, Y, R^(y), m, and p are as described herein.

In certain embodiments, Y is O. In certain embodiments, Y is S.

As defined generally above, R¹ is hydrogen, R^(z), or —C(O)R^(z),wherein R^(z) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R¹ is hydrogen. In some embodiments, R¹ is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is unsubstituted C₁₋₆alkyl. In certain embodiments, R¹ is methyl, ethyl, or propyl. In someembodiments, R¹ is —C(O)R^(z), wherein R^(z) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, R¹ is —C(O)R^(z), wherein R^(z) isunsubstituted C₁₋₆ alkyl. In certain embodiments, R¹ is acetyl.

As defined generally above, G is NR², CR³R⁴, O or S. In certainembodiments, G is NR². In certain embodiments, G is CR³R⁴. In certainembodiments, G is O. In certain embodiments, G is S.

As defined generally above, R² is selected from the group consisting ofoptionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted aryl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —C(O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A),—C(═S)N(R^(B))₂, —S(═O)R^(A), —SO₂R^(A), and —SO₂N(R^(B))₂. In someembodiments, R² is optionally substituted aryl. In certain embodiments,R² is optionally substituted phenyl. In certain embodiments, R² isunsubstituted phenyl. In certain embodiments, R² is halophenyl. Incertain embodiments, R² is fluorophenyl. In certain embodiments, R² ischlorophenyl. In some embodiments, R² is phenyl substituted withoptionally substituted C₁₋₆ alkyl. In some embodiments, R² is phenylsubstituted with optionally substituted C₁₋₃ alkyl. In certainembodiments, R² is phenyl substituted with methyl. In certainembodiments, R² is phenyl substituted with —CH₂OOH. In some embodiments,R² is phenyl substituted with a heterocyclic ring. In certainembodiments, R² is phenyl substituted with morpholinyl. In certainembodiments, R² is phenyl substituted with tetrahydropyranyl. In someembodiments, R² is optionally substituted heteroaryl. In certainembodiments, R² is optionally substituted quinoline. In certainembodiments, R² is unsubstituted quinoline. In certain embodiments, R²is substituted quinoline. In certain embodiments, R² is optionallysubstituted pyridine. In certain embodiments, R² is pyridine substitutedwith a heterocyclic ring. In some embodiments, R² is optionallysubstituted aliphatic. In certain embodiments, R² is unsubstitutedaliphatic. In certain embodiments, R² is —CH₂-aryl. In certainembodiments, R² is benzyl. In certain embodiments, R² is—CH₂-heteroaryl. In certain embodiments, R² is —CH₂-pyridyl. In someembodiments, R² is —C(═O)R^(A). In certain embodiments, R² is—C(═O)R^(A), wherein R^(A) is optionally substituted aliphatic. Incertain embodiments, R² is acetyl. In certain embodiments, R² is—SO₂R^(A). In certain embodiments, R² is —SO₂R^(A), wherein R^(A) isoptionally substituted aliphatic. In certain embodiments, R² is —SO₂CH₃.

In certain embodiments, R² is selected from the group consisting of:

As defined generally above, R³ is selected from the group consisting ofhydrogen, halo, optionally substituted aliphatic, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂. In some embodiments, R³ is selectedfrom the group consisting of hydrogen, halo, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(═O)R^(A),—NR^(B)C(═O)N(R^(B))₂, —SC(═O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(═O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂.

In certain embodiments, R³ is hydrogen. In some embodiments, R³ is nothydrogen. In some embodiments, R³ is halo. In certain embodiments, R³ isfluoro. In some embodiments, R³ is optionally substituted aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ aliphatic. Incertain embodiments, R³ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R³ is substituted C₁₋₆ alkyl. In certain embodiments, R³ is—CF₃, —CHF₂, or —CH₂F. In certain embodiments, R³ is unsubstituted C₁₋₆alkyl. In certain embodiments, R³ is methyl, ethyl, or propyl. In someembodiments, R³ is —CN or —NO₂. In some embodiments, R³ is optionallysubstituted carbocyclyl, optionally substituted phenyl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl. In someembodiments, R is —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —OC(O)R^(A), —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —SC(O)R^(A), —C(═NR^(B))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂. In some embodiments, R³ is optionallysubstituted aryl. In certain embodiments, R³ is optionally substitutedphenyl. In certain embodiments, R³ is unsubstituted phenyl. In certainembodiments, R³ is halophenyl. In certain embodiments, R³ isfluorophenyl. In certain embodiments, R³ is chlorophenyl. In someembodiments, R³ is phenyl substituted with optionally substituted C₁₋₆alkyl. In some embodiments, R³ is phenyl substituted with optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R³ is phenyl substitutedwith methyl. In certain embodiments, R³ is phenyl substituted with—CH₂OH. In some embodiments, R³ is phenyl substituted with aheterocyclic ring. In certain embodiments, R³ is phenyl substituted withmorpholinyl. In certain embodiments, R³ is phenyl substituted withtetrahydropyranyl. In some embodiments, R³ is optionally substitutedheteroaryl. In certain embodiments, R³ is optionally substitutedquinoline. In certain embodiments, R³ is unsubstituted quinoline. Incertain embodiments, R³ is substituted quinoline. In certainembodiments, R³ is optionally substituted pyridine. In certainembodiments, R³ is pyridine substituted with a heterocyclic ring. Insome embodiments, R³ is optionally substituted aliphatic. In certainembodiments, R³ is unsubstituted aliphatic. In certain embodiments, R³is —CH₂-aryl. In certain embodiments, R³ is benzyl. In certainembodiments, R³ is —CH₂-heteroaryl. In certain embodiments, R³ is—CH₂-pyridyl.

As defined generally above, R⁴ is selected from the group consisting ofhydrogen, halo, and optionally substituted aliphatic. In certainembodiments, R⁴ is hydrogen. In some embodiments, R⁴ is not hydrogen. Insome embodiments, R⁴ is halo. In certain embodiments, R⁴ is fluoro. Insome embodiments, R⁴ is optionally substituted aliphatic. In certainembodiments, R⁴ is optionally substituted C₁₋₆ aliphatic. In certainembodiments, R⁴ is optionally substituted C₁₋₆ alkyl. In certainembodiments, R⁴ is substituted C₁₋₆ alkyl. In certain embodiments, R⁴ isunsubstituted C₁₋₆ alkyl. In certain embodiments, R⁴ is methyl, ethyl,or propyl.

As defined generally above, R⁵, R⁶, R⁷, and R⁸ are each independentlyhydrogen, halo, or optionally substituted aliphatic. In someembodiments, R⁵, R⁶, R⁷, and R⁸ are hydrogen. In some embodiments, R⁶,R⁷, and R⁸ are hydrogen, and R⁵ is optionally substituted aliphatic. Insome embodiments, R⁶, R⁷, and R⁸ are hydrogen, and R⁵ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁶, R⁷, and R⁸ arehydrogen, and R⁵ is optionally substituted C₁₋₃ aliphatic. In someembodiments, R⁶, R⁷, and R⁸ are hydrogen, and R⁵ is methyl. In someembodiments, R⁶, R⁷, and R⁵ are hydrogen, and R is optionallysubstituted aliphatic. In some embodiments, R⁶, R⁷, and R⁵ are hydrogen,and R⁸ is optionally substituted C₁₋₆ aliphatic. In some embodiments,R⁶, R⁷, and R⁵ are hydrogen, and R⁸ is optionally substituted C₁₋₃aliphatic. In some embodiments, R⁶, R⁷, and R⁵ are hydrogen, and R⁸ ismethyl. In some embodiments, R⁵ is hydrogen. In some embodiments, R⁵ ishalo. In certain embodiments, R⁵ is fluoro. In some embodiments, R⁵ isoptionally substituted C₁₋₆ aliphatic. In some embodiments, R⁵ isoptionally substituted C₁₋₃ alkyl. In certain embodiments, R⁵ is methyl.In some embodiments, R⁶ is hydrogen. In some embodiments, R⁶ is halo. Incertain embodiments, R⁶ is fluoro. In some embodiments, R⁶ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁶ is optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R⁶ is methyl. In someembodiments, R⁷ is hydrogen. In some embodiments, R⁷ is halo. In certainembodiments, R⁷ is fluoro. In some embodiments, R⁷ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁷ is optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R⁷ is methyl. In someembodiments, R⁸ is hydrogen. In some embodiments, R⁸ is halo. In certainembodiments, R⁸ is fluoro. In some embodiments, R⁸ is optionallysubstituted C₁₋₆ aliphatic. In some embodiments, R⁸ is optionallysubstituted C₁₋₃ alkyl. In certain embodiments, R⁸ is methyl.

As defined generally above, p is 0, 1, or 2. In certain embodiments, pis 0. In certain embodiments, p is 1. In certain embodiments, p is 2.

As defined generally above, each R^(y) is independently selected fromthe group consisting of halo, —CN, —NO₂, optionally substitutedaliphatic, optionally substituted carbocyclyl, optionally substitutedaryl, optionally substituted heterocyclyl, optionally substitutedheteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A),—C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A),—OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂, wherein R^(A) and R^(B) aredescribed herein.

In some embodiments, at least one R^(y) is halo. In certain embodiments,at least one R^(y) is fluoro. In certain embodiments, at least one R^(y)is chloro. In some embodiments, at least one R^(y) is —CN. In someembodiments, at least one R^(y) is —OR^(A), wherein R^(A) is optionallysubstituted aliphatic. In some embodiments, at least one R^(y) is—OR^(A), wherein R^(A) is unsubstituted C₁₋₆ alkyl. In certainembodiments, at least one R^(y) is methoxy, ethoxy, or propoxy. Incertain embodiments, at least one R^(y) is methoxy. In some embodiments,at least one R^(y) is —OR^(A), wherein R^(A) is substituted C₁₋₆ alkyl.In certain embodiments, at least one R^(y) is —OCH₂CH₂N(CH₃)₂. In someembodiments, at least one R^(y) is —OR^(A), wherein R^(A) is optionallysubstituted heterocyclyl. In some embodiments, at least one R^(y) is—OR^(A), wherein R^(A) is an optionally substituted 4- to 7-memberedheterocyclyl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, at least one R^(y) is—OR^(A), wherein R^(A) is oxetanyl, tetrahydrofuranyl, ortetrahydropyranyl. In some embodiments, at least one R^(y) is—N(R^(B))₂. In some embodiments, at least one R^(y) is —N(R^(B))₂,wherein each R^(B) is independently hydrogen or C₁₋₆ alkyl. In someembodiments, at least one R^(y) is —NHR^(B). In some embodiments, atleast one R^(y) is —N(C₁₋₆ alkyl)₂, —NH(C₁₋₆ alkyl), or —NH₂. In certainembodiments, at least one R^(y) is —NH₂. In certain embodiments, atleast one R^(y) is —NHCH₃. In certain embodiments, at least one R^(y) is—N(CH₃)₂. In some embodiments, at least one R^(y) is —N(R^(B))₂ or—NHR^(B), wherein at least one R^(B) is optionally substitutedheterocyclyl. In some embodiments, at least one R^(y) is —N(R^(B))₂ or—NHR^(B), wherein at least one R^(B) is an optionally substituted 4- to7-membered heterocyclyl having 1-2 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. In some embodiments, at least oneR^(y) is —N(R^(B))₂ or —NHR^(B), wherein at least one R^(B) is oxetanyl,tetrahydropyranyl, or tetrahydrofuranyl. In some embodiments, at leastone R^(y) is —N(R^(B))₂ or —NHR^(B), wherein at least one R^(B) isoptionally substituted piperidinyl or optionally substitutedpiperazinyl.

In some embodiments, at least one R^(y) is optionally substitutedaliphatic. In certain embodiments, at least one R^(y) is substitutedaliphatic. In certain embodiments, at least one R^(y) is unsubstitutedaliphatic. In some embodiments, at least one R^(y) is optionallysubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) isunsubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) issubstituted C₁₋₆ alkyl. In certain embodiments, at least one R^(y) ismethyl, ethyl, or propyl. In certain embodiments, at least one R^(y) ismethyl. In certain embodiments, at least one R^(y) is —CF₃, CHF₂, orCH₂F. In certain embodiments, at least one R^(y) is C₁₋₆ alkylsubstituted with aryl, heteroaryl, or heterocyclyl. In certainembodiments, at least one R^(y) is benzyl. In certain embodiments, atleast one R^(y) is —(C₁₋₆ alkyl)-aryl. In certain embodiments, at leastone R^(y) is —(C₁₋₆ alkyl)-heteroaryl. In certain embodiments, at leastone R^(y) is —(C₁₋₆ alkyl)-heterocyclyl. In certain embodiments, atleast one R^(y) is —CH₂-aryl. In certain embodiments, at least one R^(y)is —CH₂-heteroaryl. In certain embodiments, at least one R^(y) is—CH₂-heterocyclyl.

In some embodiments, at least one R^(y) is —C(O)N(R^(B))₂. In certainembodiments, at least one R^(y) is —C(O)NHR^(B). In certain embodiments,at least one R^(y) is —C(O)NH₂. In certain embodiments, at least oneR^(y) is —C(O)N(R^(B))₂, wherein the R^(B) groups are taken togetherwith their intervening atoms to form an optionally substituted 5- to6-membered heterocyclyl. In certain embodiments, at least one R^(y) is—C(O)N(R^(B))₂, wherein the R^(B) groups are taken together with theirintervening atoms to form an optionally substituted morpholinyl.

In some embodiments, at least one R^(y) is —SO₂N(R^(B))₂. In certainembodiments, at least one R^(y) is —SO₂NHR^(B). In certain embodiments,at least one R^(y) is —SO₂NH₂. In certain embodiments, at least oneR^(y) is —SO₂N(R^(B))₂, wherein neither R^(B) is hydrogen. In certainembodiments, at least one R^(y) is —SO₂NH(C₁₋₆ alkyl) or —SO₂N(C₁₋₆alkyl)₂. In certain embodiments, at least one R^(y) is —SO₂N(CH₃)₂. Incertain embodiments, at least one R^(y) is —SO₂N(R^(B))₂, wherein theR^(B) groups are taken together with their intervening atoms to form anoptionally substituted 5- to 6-membered heterocyclyl. In certainembodiments, at least one R^(y) is —SO₂-morpholinyl. In certainembodiments, at least one R^(y) is —SO₂-piperidinyl, —SO₂-piperazinyl,or —SO₂-piperidinyl.

In some embodiments, at least one R^(y) is —SO₂R^(A). In someembodiments, at least one R^(y) is —SO₂R^(A), wherein R^(A) isoptionally substituted aliphatic. In some embodiments, at least oneR^(y) is —SO₂(C₁₋₆ alkyl). In some embodiments, at least one R^(y) is—SO₂CH₃. In some embodiments, at least one R^(y) is —C(O)R^(A). In someembodiments, at least one R^(y) is —C(O)R^(A), wherein R^(A) isoptionally substituted aliphatic. In some embodiments, at least oneR^(y) is —C(O)(C₁₋₆ alkyl). In some embodiments, at least one R^(y) is—C(O)CH₃.

In some embodiments, at least one R^(y) is —N(R^(B))C(O)R^(A). Incertain embodiments, at least one R^(y) is —NHC(O)R^(A). In certainembodiments, at least one R^(y) is —NHC(O)(C₁₋₆ alkyl). In certainembodiments, at least one R^(y) is —NHC(O)CH₃.

In some embodiments, at least one R^(y) is —N(R^(B))SO₂R^(A). In someembodiments, at least one R^(y) is —NHSO₂R^(A). In some embodiments, atleast one R^(y) is —N(C₁₋₆ alkyl)SO₂R^(A). In certain embodiments, atleast one R^(y) is —NHSO₂(C₁₋₆ alkyl) or —N(C₁₋₆ alkyl)SO₂(C₁₋₆ alkyl).In certain embodiments, at least one R^(y) is —NHSO₂CH₃. In certainembodiments, at least one R^(y) is —N(CH₃)SO₂CH₃.

In some embodiments, at least one R^(y) is optionally substitutedheterocyclyl, optionally substituted carbocyclyl, optionally substitutedaryl, or optionally substituted heteroaryl. In certain embodiments, atleast one R^(y) is an optionally substituted 5- to 6-memberedheterocyclyl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is an optionally substituted 5-membered heterocyclyl having oneheteroatom selected from nitrogen, oxygen, and sulfur. In certainembodiments, at least one R^(y) is optionally substituted pyrrolidinyl.In certain embodiments, at least one R^(y) is pyrroldinyl,hydroxypyrrolidinyl, or methylpyrrolidinyl. In certain embodiments, atleast one R^(y) is an optionally substituted 6-membered heterocyclylhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, at least one R^(y) is an optionallysubstituted 6-membered heterocyclyl having one heteroatom selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is optionally substituted piperidinyl. In certain embodiments, at leastone R^(y) is an optionally substituted 6-membered heterocyclyl havingtwo heteroatoms independently selected from nitrogen, oxygen, andsulfur. In certain embodiments, at least one R^(y) is optionallysubstituted piperdinyl, optionally substituted piperazinyl, oroptionally substituted morpholinyl. In certain embodiments, at least oneR^(y) is morpholinyl, tetrahydropyranyl, piperidinyl, methylpiperidinyl,piperazinyl, methylpiperazinyl, acetylpiperazinyl,methylsulfonylpiperazinyl, aziridinyl, or methylaziridinyl. In someembodiments, at least one R^(y) is an optionally substituted 5- to6-membered heteroaryl having 1-3 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, at least one R^(y)is an optionally substituted 5-membered heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur. Incertain embodiments, at least one R^(y) is an optionally substituted5-membered heteroaryl having one heteroatom selected from nitrogen,oxygen, and sulfur. In certain embodiments, at least one R^(y) is anoptionally substituted 5-membered heteroaryl having two heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In certainembodiments, at least one R^(y) is an optionally substituted 6-memberedheteroaryl having 1-3 nitrogens. In certain embodiments, at least oneR^(y) is an optionally substituted pyrazolyl. In certain embodiments, atleast one R^(y) is an optionally substituted imidazolyl. In certainembodiments, at least one R^(y) is an optionally substituted pyridyl. Incertain embodiments, at least one R^(y) is an optionally substitutedpyrimidyl. In certain embodiments, at least one R^(y) is pyrazolyl,methylpyrazolyl, imidazolyl, or methylimidazolyl.

In some embodiments, R^(y) is —OR^(A). In some embodiments, R^(y) is—OR^(A), wherein R^(A) is optionally substituted heterocyclyl. In someembodiments, R^(y) is —OR^(A), wherein R^(A) is optionally substitutedheteroaryl. In some embodiments, R^(y) is —OR^(A), wherein R^(A) isoptionally substituted cycloalkyl. In some embodiments, R^(y) is—N(R^(B))₂. In some embodiments, R^(y) is —NHR^(B). In some embodiments,R^(y) is —NHR^(B), wherein R^(B) is optionally substituted heterocyclyl.In some embodiments, R^(y) is —NHR^(B), wherein R^(B) is optionallysubstituted heteroaryl. In some embodiments, R^(y) is —NHR^(B), whereinR^(B) is optionally substituted cycloalkyl. In some embodiments, R^(y)is —N(R^(B))₂, wherein one R^(B) is optionally substituted heterocyclyl,and the other R^(B) s C₁₋₄ alkyl. In some embodiments, R^(y) is—N(R^(B))₂, wherein one R^(B) is optionally substituted heteroaryl, andthe other R^(B) is C₁₋₄ alkyl. In some embodiments, R^(y) is —N(R^(B))₂,wherein one R^(B) is optionally substituted cycloalkyl, and the otherR^(B) is C₁₋₄ alkyl.

As defined generally above, each R^(x) is independently selected fromthe group consisting of halo, —CN, optionally substituted aliphatic,—OR′, and —N(R″)₂. In certain embodiments, at least one R^(x) is halo.In certain embodiments, at least one R^(x) is fluoro. In certainembodiments, at least one R^(x) is —CN. In certain embodiments, at leastone R^(x) is optionally substituted aliphatic. In certain embodiments,at least one R^(x) is optionally substituted C₁₋₆ alkyl. In certainembodiments, at least one R^(x) is methyl. In certain embodiments, atleast one R^(x) is —CF₃. In certain embodiments, at least one R^(x) is—OR′ or —N(R″)₂. In certain embodiments, R^(x) is not —OR′ or —N(R″)₂.In certain embodiments, at least one R^(x) is —OCH₃. In certainembodiments, R^(x) is not —OCH₃.

As is generally understood from the above disclosure, the ring system:

is a fused bicyclic ring system, i.e., a phenyl ring fused to a nitrogencontaining ring, wherein the point of attachment to the parent moiety ison the nitrogen, and wherein the fused bicyclic system is optionallysubstituted with (R^(x))_(n), wherein n and R^(x) are as defined herein.As is generally understood, each of the atoms of the phenyl ring and thenitrogen-containing ring can be independently optionally substitutedwith R^(x), as valency permits.

In certain embodiments, the fused bicyclic ring system is optionallysubstituted with one or more R^(x), with the proviso that when thenitrogen-containing ring is substituted at one of the positions alpha tothe nitrogen, R^(x) is not —C(═O)R^(x1), wherein R^(x1) is optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, or —SR^(A), wherein R^(A)and R^(B) are as generally defined herein. In certain embodiments, thenitrogen-containing ring does not comprise an R^(x) substituent. Incertain embodiments, only atoms of the phenyl ring are optionallysubstituted with one or more R^(x).

In certain embodiments, the nitrogen-containing ring is optionallysubstituted and the fused bicyclic ring system is of the formula:

wherein R^(x) is as defined above, and n is 0, 1, 2, 3, or 4.

Thus, one of ordinary skill in the art will appreciate that an R^(x)group can be attached anywhere on the tetrahydroisoquinoline ordihydroisoquinoline ring. In certain embodiments, an R^(x) group isattached to the benzene portion of the tetrahydroisoquinoline ordihydroisoquinoline ring. In certain embodiments, an R^(x) group isattached to the tetrahydropyridine or dihydropyridine portion of thetetrahydroisoquinoline or dihydroisoquinoline ring. In certainembodiments, R^(x) groups are attached to both the benzene portion andthe tetrahydropyridine (or dihydropyridine) portion of thetetrahydroisoquinoline (or dihydroisoquinoline) ring. See, for example,the structures shown below:

In certain embodiments, a provided compound is of Formula (VII):

or a pharmaceutically acceptable salt thereof.

As defined generally above, n1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.In certain embodiments, n is 0. In certain embodiments, n is 1. Incertain embodiments, n is 2.

As generally defined above, R^(A1) and R^(A2) are independentlyhydrogen, substituted or unsubstituted C₁₋₃ alkyl, substituted orunsubstituted acyl, or a nitrogen protecting group. In some embodiments,R^(A1) is hydrogen. In some embodiments, R^(A1) is substituted orunsubstituted C₁₋₃ alkyl. In some embodiments, R^(A1) is unsubstitutedC₁₋₃ alkyl. In some embodiments, R^(A1) is methyl, ethyl, n-propyl, orisopropyl. In some embodiments, R^(A1) is substituted C₁₋₃ alkyl. Insome embodiments, R^(A1) is —CF₃, —CHF₂, —CH₂F, or —CH(CF₃)CH₃. In someembodiments, R^(A1) is substituted or unsubstituted acyl. In someembodiments, R^(A1) is acetyl. In some embodiments, R^(A1) is a nitrogenprotecting group. In some embodiments, R^(A1) is CH₃SO₂—. In someembodiments, R^(A2) is hydrogen. In some embodiments, R^(A2) issubstituted or unsubstituted C₁₋₃ alkyl. In some embodiments, R^(A2) isunsubstituted C₁₋₃ alkyl. In some embodiments, R^(A2) is methyl, ethyl,n-propyl, or isopropyl. In some embodiments, R^(A2) is substituted C₁₋₃alkyl. In some embodiments, R^(A2) is —CF₃, —CHF₂, —CH₂F, or—CH(CF₃)CH₃. In some embodiments, R^(A2) is substituted or unsubstitutedacyl. In some embodiments, R^(A2) is acetyl. In some embodiments, R^(A2)is a nitrogen protecting group. In some embodiments, R^(A2) is CH₃SO₂—.In some embodiments, R^(A1) is hydrogen, and R^(A2) is hydrogen. In someembodiments, R^(A1) is hydrogen, and R^(A2) is substituted orunsubstituted C₁₋₃ alkyl. In some embodiments, R^(A1) is hydrogen, andR^(A2) is methyl, ethyl, n-propyl, or isopropyl. In some embodiments,R^(A1) is hydrogen, and R^(A2) is —CF₃, —CHF₂, —CH₂F, or —CH(CF₃)CH₃. Insome embodiments, R^(A1) is hydrogen, and R^(A2) is substituted orunsubstituted acyl. In some embodiments, R^(A1) is hydrogen, and R^(A2)is acetyl. In some embodiments, R^(A1) is hydrogen, and R^(A2) is anitrogen protecting group. In some embodiments, R^(A1) is hydrogen andR^(A2) is CH₃SO₂—. In some embodiments, R^(A1) is substituted orunsubstituted C₁₋₃ alkyl, and R^(A2) is substituted or unsubstitutedC₁₋₃ alkyl. In some embodiments, R^(A1) is substituted or unsubstitutedC₁₋₃ alkyl, and R^(A2) is methyl. In some embodiments, R^(A1) issubstituted or unsubstituted C₁₋₃ alkyl, and R^(A2) is ethyl. In someembodiments, R^(A1) is substituted or unsubstituted C₁₋₃ alkyl, andR^(A2) is n-propyl. In some embodiments, R^(A1) is substituted orunsubstituted C₁₋₃ alkyl, and R^(A2) is isopropyl. In some embodiments,R^(A1) is substituted or unsubstituted C₁₋₃ alkyl, and R^(A2) issubstituted or unsubstituted acyl. In some embodiments, R^(A1) issubstituted or unsubstituted C₁₋₃ alkyl, and R^(A2) is a nitrogenprotecting group. In some embodiments, R^(A1) is methyl, and R^(A2) issubstituted or unsubstituted C₁₋₃ alkyl. In some embodiments, R^(A1) ismethyl, and R^(A2) is methyl. In some embodiments, R^(A1) is methyl, andR^(A2) is ethyl. In some embodiments, R^(A1) is methyl, and R^(A2) isn-propyl. In some embodiments, R^(A1) is methyl, and R^(A2) isisopropyl. In some embodiments, R^(A1) is methyl, and R^(A2) issubstituted or unsubstituted acyl. In some embodiments, R^(A1) ismethyl, and R^(A2) is a nitrogen protecting group. In some embodiments,R^(A1) is ethyl, and R^(A2) is substituted or unsubstituted C₁₋₃ alkyl.In some embodiments, R^(A1) is ethyl, and R^(A2) is methyl. In someembodiments, R^(A1) is ethyl, and R^(A2) is ethyl. In some embodiments,R^(A1) is ethyl, and R^(A2) is n-propyl. In some embodiments, R^(A1) isethyl, and R^(A2) is isopropyl. In some embodiments, R^(A1) is ethyl,and R^(A2) is substituted or unsubstituted acyl. In some embodiments,R^(A1) is ethyl, and R^(A2) is a nitrogen protecting group. In someembodiments, R^(A1) is n-propyl, and R^(A2) is substituted orunsubstituted C₁₋₃ alkyl. In some embodiments, R^(A1) is n-propyl, andR^(A2) is methyl. In some embodiments, R^(A1) is n-propyl, and R^(A2) isethyl. In some embodiments, R^(A1) is n-propyl, and R^(A2) is n-propyl.In some embodiments, R^(A1) is n-propyl and R^(A2) is isopropyl. In someembodiments, R^(A1) is n-propyl, and R^(A2) is substituted orunsubstituted acyl. In some embodiments, R^(A1) is n-propyl and R^(A2)is a nitrogen protecting group. In some embodiments, R^(A1) is isopropyland R^(A2) is substituted or unsubstituted C₁₋₃ alkyl. In someembodiments, R^(A1) is isopropyl and R^(A2) is methyl. In someembodiments, R^(A1) is isopropyl and R^(A2) is ethyl. In someembodiments, R^(A1) is isopropyl, and R^(A2) is n-propyl. In someembodiments, R^(A1) is isopropyl, and R^(A2) is isopropyl. In someembodiments, R^(A1) is isopropyl, and R^(A2) is substituted orunsubstituted acyl. In some embodiments, R^(A1) is isopropyl, and R^(A2)is a nitrogen protecting group. In some embodiments, R^(A1) issubstituted or unsubstituted acyl, and R^(A2) is substituted orunsubstituted C₁₋₃ alkyl. In some embodiments, R^(A1) is a nitrogenprotecting group, and R^(A2) is substituted or unsubstituted C₁₋₃ alkyl.In some embodiments, R^(A1) is a nitrogen protecting group and R^(A2) ismethyl. In some embodiments, R^(A1) is a nitrogen protecting group, andR^(A2) is ethyl. In some embodiments, R^(A1) is a nitrogen protectinggroup, and R^(A2) is n-propyl. In some embodiments, R^(A1) is a nitrogenprotecting group, and R^(A2) is isopropyl. In some embodiments, R^(A1)is a nitrogen protecting group, and R^(A2) is a nitrogen protectinggroup.

As generally defined above, R^(A1) and R^(A2) can be taken together withthe intervening nitrogen atom to form a substituted or unsubstituted 3-6membered heterocyclic ring. In certain embodiments, R^(A1) and R^(A2)can be taken together with the intervening nitrogen atom to form asubstituted or unsubstituted azetidine. In certain embodiments, R^(A1)and R^(A2) can be taken together with the intervening nitrogen atom toform a substituted or unsubstituted pyrrolidine. In certain embodiments,R^(A1) and R^(A2) can be taken together with the intervening nitrogenatom to form a substituted or unsubstituted piperidine. In certainembodiments, R^(A1) and R^(A2) can be taken together with theintervening nitrogen atom to form a substituted or unsubstitutedpiperazine. In certain embodiments, R^(A1) and R^(A2) can be takentogether with the intervening nitrogen atom to form a substituted orunsubstituted morpholine.

In some embodiments, e.g. for Formula (A), Formula (I), or any subgenerathereof, the provided compound is of a free base form. In someembodiments, e.g. for Formula (A), Formula (I), or any subgenerathereof, the provided compound is in the form of a pharmaceuticallyacceptable salt as generally defined herein. In some embodiments, theprovided compound is a hydrochloride salt thereof. In some embodiments,the provided compound is a tartrate salt thereof. In some embodiments,the provided compound is a monotartrate salt thereof. In someembodiments, the provided compound is a bitartrate salt thereof.

In certain embodiments, a provided compound is a compound listed inTable 1A, or a pharmaceutically acceptable salt thereof.

TABLE 1A Exemplary Compounds Cmpd LCMS m/z No Structure Exact Mass (M +H)  1

322.1681 323.1  2

284.1525 285.1  3

379.226  380.2  4

416.2212 417.2  5

399.1714 400.1  6

383.2009 384.2  7

399.1714 400.1  8

451.2583 452.3  9

399.1714 400.1 10

379.226  380.2 11

383.2009 384.2 12

450.2631 451.2 13

360.1838 361.2 14

360.1838 361.2 15

379.226  380.2 16

364.2151 365.2 17

364.2151 365.2 18

375.1947 376.2 19

375.1947 376.2 20

395.2209 396.2 21

331.1896 332.1 22

367.1566 368.1 23

373.2365 374.2 24

379.226  380.1 25

380.2212 381.2 26

380.2212 381.1 27

303.1947 304.1 28

290.163  291.1 29

408.2525 409.3 30

408.2525 407.2 31

338.163  339.2 32

401.2315 402.2 33

411.2158 412.1 34

412.2111 413.1

In certain embodiments, a provided compound is a compound listed inTable 1B, or a pharmaceutically acceptable salt thereof.

TABLE 1B Exemplary Compounds Cmpd LCMS m/z No Structure Exact Mass (M +H) 35

449.2791 — 36

463.2947 — 37

477.3104 — 38

491.2896 — 39

527.2566 — 40

502.2556 — 41

545.2978 — 42

517.3417 — 43

518.3369 — 44

519.3209 — 45

489.3104 — 46

503.3260 —

In certain embodiments, a provided compound inhibits PRMT5. In certainembodiments, a provided compound inhibits wild-type PRMT5. In certainembodiments, a provided compound inhibits a mutant PRMT5. In certainembodiments, a provided compound inhibits PRMT5, e.g., as measured in anassay described herein. In certain embodiments, the PRMT5 is from ahuman. In certain embodiments, a provided compound inhibits PRMT5 at anIC₅₀ less than or equal to 10 μM. In certain embodiments, a providedcompound inhibits PRMT5 at an IC₅₀ less than or equal to 1 μM. Incertain embodiments, a provided compound inhibits PRMT5 at an IC₅₀ lessthan or equal to 0.1 μM. In certain embodiments, a provided compoundinhibits PRMT5 in a cell at an EC₅₀ less than or equal to 10 μM. Incertain embodiments, a provided compound inhibits PRMT5 in a cell at anEC₅₀ less than or equal to 1 μM. In certain embodiments, a providedcompound inhibits PRMT5 in a cell at an EC₅₀ less than or equal to 0.1μM. In certain embodiments, a provided compound inhibits cellproliferation at an EC₅₀ less than or equal to 10 μM. In certainembodiments, a provided compound inhibits cell proliferation at an EC₅₀less than or equal to 1 μM. In certain embodiments, a provided compoundinhibits cell proliferation at an EC₅₀ less than or equal to 0.1 μM. Insome embodiments, a provided compound is selective for PRMT5 over othermethyltransferases. In certain embodiments, a provided compound is atleast about 10-fold selective, at least about 20-fold selective, atleast about 30-fold selective, at least about 40-fold selective, atleast about 50-fold selective, at least about 60-fold selective, atleast about 70-fold selective, at least about 80-fold selective, atleast about 90-fold selective, or at least about 100-fold selective forPRMT5 relative to one or more other methyltransferases.

It will be understood by one of ordinary skill in the art that the PRMT5can be wild-type PRMT5, or any mutant or variant of PRMT5.

In some embodiments embodiment, the mutant or variant of PRMT5 containsone or more mutations (e.g., conservative substitutions). In someembodiments, provided herein is a PRMT5 point mutant. In someembodiments, the PRMT point mutant has an amino acid sequence that adegree of homology to the amino acid sequence of SEQ ID NO: 1 of atleast about 80%, e.g., at least about 85%, at least about 90%, at leastabout 95%, or at least about 97%. Further provided is a protein that hasa degree of homology to the amino acid sequence of SEQ ID NO: 2 of atleast about 80%, e.g., at least about 85%, at least about 90%, at leastabout 95%, or at least about 97%.

In certain embodiments, the PRMT5 is isoform A (GenBank accession no.NP006100) (SEQ ID NO.:1):

MAAMAVGGAG GSRVSSGRDL NCVPEIADTL GAVAKQGFDFLCMPVFHPRF KREFIQEPAK NRPGPQTRSD LLLSGRDWNTLIVGKLSPWI RPDSKVEKIR RNSEAAMLQE LNFGAYLGLPAFLLPLNQED NTNLARVLTN HIHTGHHSSM FWMRVPLVAPEDLRDDIIEN APTTHTEEYS GEEKTWMWWH NFRTLCDYSKRIAVALEIGA DLPSNHVIDR WLGEPIKAAI LPTSIFLTNKKGFPVLSKMH QRLIFRLLKL EVQFIITGTN HHSEKEFCSYLQYLEYLSQN RPPPNAYELF AKGYEDYLQS PLQPLMDNLESQTYEVFEKD PIKYSQYQQA IYKCLLDRVP EEEKDTNVQVLMVLGAGRGP LVNASLRAAK QADRRIKLYA VEKNPNAVVTLENWQFEEWG SQVTVVSSDM REWVAPEKAD IIVSELLGSFADNELSPECL DGAQHFLKDD GVSIPGEYTS FLAPISSSKLYNEVRACREK DRDPEAQFEM PYVVRLHNFH QLSAPQPCFTFSHPNRDPMI DNNRYCTLEF PVEVNTVLHG FAGYFETVLYQDITLSIRPE THSPGMFSWF PILFPIKQPI TVREGQTICVRFWRCSNSKK VWYEWAVTAP VCSAIHNPTG RSYTIGL

In certain embodiments, the PRMT5 is isoform B (GenBank accession no.NP001034708) (SEQ ID NO.:2)

MRGPNSGTEK GRLVIPEKQG FDFLCMPVFH PRFKREFIQEPAKNRPGPQT RSDLLLSGRD WNTLIVGKLS PWIRPDSKVEKIRRNSEAAM LQELNFGAYL GLPAFLLPLN QEDNTNLARVLTNHIHTGHH SSMFWMRVPL VAPEDLRDDI IENAPTTHTEEYSGEEKTWM WWHNFRTLCD YSKRIAVALE IGADLPSNHVIDRWLGEPIK AAILPTSIFL TNKKGFPVLS KMHQRLIFRLLKLEVQFIIT GTNHHSEKEF CSYLQYLEYL SQNRPPPNAYELFAKGYEDY LQSPLQPLMD NLESQTYEVF EKDPIKYSQYQQAIYKCLLD RVPEEEKDTN VQVLMVLGAG RGPLVNASLRAAKQADRRIK LYAVEKNPNA VVTLENWQFE EWGSQVTVVSSDMREWVAPE KADIIVSELL GSFADNELSP ECLDGAQHFLKDDGVSIPGE YTSFLAPISS SKLYNEVRAC REKDRDPEAQFEMPYVVRLH NFHQLSAPQP CFTFSHPNRD PMIDNNRYCTLEFPVEVNTV LHGFAGYFET VLYQDITLSI RPETHSPGMFSWFPILFPIK QPITVREGQT ICVRFWRCSN SKKVWYEWAV TAPVCSAIHN PTGRSYTIGL

In certain embodiments, the PRMT5 is transcript variant 1 (GenBankaccession no. NM_006109).

The present disclosure provides pharmaceutical compositions comprising acompound described herein, e.g., a compound of Formula (A), e.g.,Formula (I), or a pharmaceutically acceptable salt thereof, as describedherein, and optionally a pharmaceutically acceptable excipient. It willbe understood by one of ordinary skill in the art that the compoundsdescribed herein, or salts thereof, may be present in various forms,such as amorphous, hydrates, solvates, or polymorphs. In certainembodiments, a provided composition comprises two or more compoundsdescribed herein. In certain embodiments, a compound described herein,or a pharmaceutically acceptable salt thereof, is provided in aneffective amount in the pharmaceutical composition. In certainembodiments, the effective amount is a therapeutically effective amount.In certain embodiments, the effective amount is an amount effective forinhibiting PRMT5. In certain embodiments, the effective amount is anamount effective for treating a PRMT5-mediated disorder. In certainembodiments, the effective amount is a prophylactically effectiveamount. In certain embodiments, the effective amount is an amounteffective to prevent a PRMT5-mediated disorder.

In certain embodiments, the provided pharmaceutical compositionscomprise a compound described herein, e.g., a compound of Formula (A),e.g., Formula (I), or any subgenera thereof, and optionally apharmaceutically acceptable excipient, wherein the compound is of a freebase form. In certain embodiments, the provided pharmaceuticalcompositions comprise a compound described herein, e.g., a compound ofFormula (A), e.g., Formula (I), or any subgenera thereof, and optionallya pharmaceutically acceptable excipient, wherein the compound is in theform of a pharmaceutically acceptable salt as generally defined herein.In certain embodiments, the provided pharmaceutical compositionscomprise a hydrochloride salt of a compound described herein andoptionally a pharmaceutically acceptable excipient. In certainembodiments, the provided pharmaceutical compositions comprise atartrate salt of a compound described herein and optionally apharmaceutically acceptable excipient. In certain embodiments, theprovided pharmaceutical compositions comprise a monotartrate salt of acompound described herein and optionally a pharmaceutically acceptableexcipient. In certain embodiments, the provided pharmaceuticalcompositions comprise a bitartrate salt of a compound described hereinand optionally a pharmaceutically acceptable excipient. In certainembodiments, the provided pharmaceutical compositions comprise amonotartrate salt and a bitartrate salt of a compound described hereinand optionally a pharmaceutically acceptable excipient. In certainembodiments, the provided pharmaceutical compositions comprise acompound described herein in a form of free base, and a pharmaceuticallyacceptable salt thereof, and optionally a pharmaceutically acceptableexcipient.

Pharmaceutically acceptable excipients include any and all solvents,diluents, or other liquid vehicles, dispersions, suspension aids,surface active agents, isotonic agents, thickening or emulsifyingagents, preservatives, solid binders, lubricants, and the like, assuited to the particular dosage form desired. General considerations informulation and/or manufacture of pharmaceutical compositions agents canbe found, for example, in Remington's Pharmaceutical Sciences, SixteenthEdition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980), andRemington: The Science and Practice of Pharmacy, 21st Edition(Lippincott Williams & Wilkins, 2005).

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include the steps of bringing a compound described herein (the“active ingredient”) into association with a carrier and/or one or moreother accessory ingredients, and then, if necessary and/or desirable,shaping and/or packaging the product into a desired single- ormulti-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.As used herein, a “unit dose” is discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient is generally equal to the dosage ofthe active ingredient which would be administered to a subject and/or aconvenient fraction of such a dosage such as, for example, one-half orone-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition of the present disclosure will vary,depending upon the identity, size, and/or condition of the subjecttreated and further depending upon the route by which the composition isto be administered. By way of example, the composition may comprisebetween 0.1% and 100% (w/w) active ingredient.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

Exemplary diluents include calcium carbonate, sodium carbonate, calciumphosphate, dicalcium phosphate, calcium sulfate, calcium hydrogenphosphate, sodium phosphate lactose, sucrose, cellulose,microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodiumchloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

Exemplary granulating and/or dispersing agents include potato starch,corn starch, tapioca starch, sodium starch glycolate, clays, alginicacid, guar gum, citrus pulp, agar, bentonite, cellulose and woodproducts, natural sponge, cation-exchange resins, calcium carbonate,silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone)(crospovidone), sodium carboxymethyl starch (sodium starch glycolate),carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose(croscarmellose), methylcellulose, pregelatinized starch (starch 1500),microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate,quaternary ammonium compounds, and mixtures thereof.

Exemplary surface active agents and/or emulsifiers include naturalemulsifiers (e.g., acacia, agar, alginic acid, sodium alginate,tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk,casein, wool fat, cholesterol, wax, and lecithin), colloidal clays(e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminumsilicate)), long chain amino acid derivatives, high molecular weightalcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetinmonostearate, ethylene glycol distearate, glyceryl monostearate, andpropylene glycol monostearate, polyvinyl alcohol), carbomers (e.g.,carboxy polymethylene, polyacrylic acid, acrylic acid polymer, andcarboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,carboxymethylcellulose sodium, powdered cellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylenesorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60),polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate(Span 40), sorbitan monostearate (Span 60], sorbitan tristearate (Span65), glyceryl monooleate, sorbitan monooleate (Span 80)),polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45),polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil,polyoxymethylene stearate, and Solutol), sucrose fatty acid esters,polyethylene glycol fatty acid esters (e.g., Cremophor™),polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij 30)),poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamineoleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 188, cetrimoniumbromide, cetylpyridinium chloride, benzalkonium chloride, docusatesodium, and/or mixtures thereof.

Exemplary binding agents include starch (e.g., cornstarch and starchpaste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin,molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums(e.g., acacia, sodium alginate, extract of Irish moss, panwar gum,ghatti gum, mucilage of isapol husks, carboxymethylcellulose,methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate(Veegum), and larch arabogalactan), alginates, polyethylene oxide,polyethylene glycol, inorganic calcium salts, silicic acid,polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

Exemplary preservatives include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, alcoholpreservatives, acidic preservatives, and other preservatives.

Exemplary antioxidants include alpha tocopherol, ascorbic acid, acorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, potassium metabisulfite, propionic acid, propylgallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, andsodium sulfite.

Exemplary chelating agents include ethylenediaminetetraacetic acid(EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodiumedetate, trisodium edetate, calcium disodium edetate, dipotassiumedetate, and the like), citric acid and salts and hydrates thereof(e.g., citric acid monohydrate), fumaric acid and salts and hydratesthereof, malic acid and salts and hydrates thereof, phosphoric acid andsalts and hydrates thereof, and tartaric acid and salts and hydratesthereof. Exemplary antimicrobial preservatives include benzalkoniumchloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide,cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol,chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,propylene glycol, and thimerosal.

Exemplary antifungal preservatives include butyl paraben, methylparaben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoicacid, potassium benzoate, potassium sorbate, sodium benzoate, sodiumpropionate, and sorbic acid.

Exemplary alcohol preservatives include ethanol, polyethylene glycol,phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate,and phenylethyl alcohol.

Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E,beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbicacid, sorbic acid, and phytic acid.

Other preservatives include tocopherol, tocopherol acetate, deteroximemesylate, cetrimide, butylated hydroxyanisol (BHA), butylatedhydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS),sodium lauryl ether sulfate (SLES), sodium bisulfite, sodiummetabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus,Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, andEuxyl. In certain embodiments, the preservative is an anti-oxidant. Inother embodiments, the preservative is a chelating agent.

Exemplary buffering agents include citrate buffer solutions, acetatebuffer solutions, phosphate buffer solutions, ammonium chloride, calciumcarbonate, calcium chloride, calcium citrate, calcium glubionate,calcium gluceptate, calcium gluconate, D-gluconic acid, calciumglycerophosphate, calcium lactate, propanoic acid, calcium levulinate,pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasiccalcium phosphate, calcium hydroxide phosphate, potassium acetate,potassium chloride, potassium gluconate, potassium mixtures, dibasicpotassium phosphate, monobasic potassium phosphate, potassium phosphatemixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodiumcitrate, sodium lactate, dibasic sodium phosphate, monobasic sodiumphosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide,aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline,Ringer's solution, ethyl alcohol, and mixtures thereof.

Exemplary lubricating agents include magnesium stearate, calciumstearate, stearic acid, silica, talc, malt, glyceryl behanate,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate,sodium lauryl sulfate, and mixtures thereof.

Exemplary natural oils include almond, apricot kernel, avocado, babassu,bergamot, black current seed, borage, cade, camomile, canola, caraway,carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee,corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed,geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate,jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademianut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, andwheat germ oils. Exemplary synthetic oils include, but are not limitedto, butyl stearate, caprylic triglyceride, capric triglyceride,cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate,mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixturesthereof.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents. Incertain embodiments for parenteral administration, the compoundsdescribed herein are mixed with solubilizing agents such as Cremophor™,alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins,polymers, and mixtures thereof.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This can be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Compositions for rectal or vaginal administration are typicallysuppositories which can be prepared by mixing the compounds describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active ingredient.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may comprise buffering agents.

Solid compositions of a similar type can be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes. Solid compositions of asimilar type can be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active ingredient can be in micro-encapsulated form with one or moreexcipients as noted above. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active ingredient can be admixed with at least oneinert diluent such as sucrose, lactose, or starch. Such dosage forms maycomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets, and pills, the dosage forms may comprise bufferingagents. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of a providedcompound may include ointments, pastes, creams, lotions, gels, powders,solutions, sprays, inhalants and/or patches. Generally, the activeingredient is admixed under sterile conditions with a pharmaceuticallyacceptable carrier and/or any desired preservatives and/or buffers ascan be required. Additionally, the present disclosure encompasses theuse of transdermal patches, which often have the added advantage ofproviding controlled delivery of an active ingredient to the body. Suchdosage forms can be prepared, for example, by dissolving and/ordispensing the active ingredient in the proper medium. Alternatively oradditionally, the rate can be controlled by either providing a ratecontrolling membrane and/or by dispersing the active ingredient in apolymer matrix and/or gel.

Suitable devices for use in delivering intradermal pharmaceuticalcompositions described herein include short needle devices such as thosedescribed in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288;4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositionscan be administered by devices which limit the effective penetrationlength of a needle into the skin, such as those described in PCTpublication WO 99/34850 and functional equivalents thereof. Jetinjection devices which deliver liquid vaccines to the dermis via aliquid jet injector and/or via a needle which pierces the stratumcorneum and produces a jet which reaches the dermis are suitable. Jetinjection devices are described, for example, in U.S. Pat. Nos.5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189;5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335;5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880;4,940,460; and PCT publications WO 97/37705 and WO 97/13537. Ballisticpowder/particle delivery devices which use compressed gas to acceleratevaccine in powder form through the outer layers of the skin to thedermis are suitable. Alternatively or additionally, conventionalsyringes can be used in the classical mantoux method of intradermaladministration.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi liquid preparations such as liniments,lotions, oil in water and/or water in oil emulsions such as creams,ointments and/or pastes, and/or solutions and/or suspensions.Topically-administrable formulations may, for example, comprise fromabout 1% to about 10% (w/w) active ingredient, although theconcentration of the active ingredient can be as high as the solubilitylimit of the active ingredient in the solvent. Formulations for topicaladministration may further comprise one or more of the additionalingredients described herein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration using a device comprising a dry powderreservoir to which a stream of propellant can be directed to dispersethe powder and/or using a self propelling solvent/powder dispensingcontainer such as a device comprising the active ingredient dissolvedand/or suspended in a low-boiling propellant in a sealed container. Suchpowders comprise particles wherein at least 98% of the particles byweight have a diameter greater than 0.5 nanometers and at least 95% ofthe particles by number have a diameter less than 7 nanometers.Alternatively, at least 95% of the particles by weight have a diametergreater than 1 nanometer and at least 90% of the particles by numberhave a diameter less than 6 nanometers. Dry powder compositions mayinclude a solid fine powder diluent such as sugar and are convenientlyprovided in a unit dose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions formulated for pulmonary delivery mayprovide the active ingredient in the form of droplets of a solutionand/or suspension. Such formulations can be prepared, packaged, and/orsold as aqueous and/or dilute alcoholic solutions and/or suspensions,optionally sterile, comprising the active ingredient, and mayconveniently be administered using any nebulization and/or atomizationdevice. Such formulations may further comprise one or more additionalingredients including, but not limited to, a flavoring agent such assaccharin sodium, a volatile oil, a buffering agent, a surface activeagent, and/or a preservative such as methylhydroxybenzoate. The dropletsprovided by this route of administration may have an average diameter inthe range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition. Anotherformulation suitable for intranasal administration is a coarse powdercomprising the active ingredient and having an average particle fromabout 0.2 to 500 micrometers. Such a formulation is administered byrapid inhalation through the nasal passage from a container of thepowder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) and as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein. A provided pharmaceutical composition can be prepared,packaged, and/or sold in a formulation for buccal administration. Suchformulations may, for example, be in the form of tablets and/or lozengesmade using conventional methods, and may contain, for example, 0.1 to20% (w/w) active ingredient, the balance comprising an orallydissolvable and/or degradable composition and, optionally, one or moreof the additional ingredients described herein. Alternately,formulations for buccal administration may comprise a powder and/or anaerosolized and/or atomized solution and/or suspension comprising theactive ingredient. Such powdered, aerosolized, and/or aerosolizedformulations, when dispersed, may have an average particle and/ordroplet size in the range from about 0.1 to about 200 nanometers, andmay further comprise one or more of the additional ingredients describedherein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation for ophthalmic administration. Such formulationsmay, for example, be in the form of eye drops including, for example, a0.1/1.0% (w/w) solution and/or suspension of the active ingredient in anaqueous or oily liquid carrier. Such drops may further comprisebuffering agents, salts, and/or one or more other of the additionalingredients described herein. Other opthalmically-administrableformulations which are useful include those which comprise the activeingredient in microcrystalline form and/or in a liposomal preparation.Ear drops and/or eye drops are contemplated as being within the scope ofthis disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

Compounds provided herein are typically formulated in dosage unit formfor ease of administration and uniformity of dosage. It will beunderstood, however, that the total daily usage of provided compositionswill be decided by the attending physician within the scope of soundmedical judgment. The specific therapeutically effective dose level forany particular subject or organism will depend upon a variety of factorsincluding the disease, disorder, or condition being treated and theseverity of the disorder; the activity of the specific active ingredientemployed; the specific composition employed; the age, body weight,general health, sex and diet of the subject; the time of administration,route of administration, and rate of excretion of the specific activeingredient employed; the duration of the treatment; drugs used incombination or coincidental with the specific active ingredientemployed; and like factors well known in the medical arts.

The compounds and compositions provided herein can be administered byany route, including enteral (e.g., oral), parenteral, intravenous,intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by powders, ointments,creams, and/or drops), mucosal, nasal, bucal, sublingual; byintratracheal instillation, bronchial instillation, and/or inhalation;and/or as an oral spray, nasal spray, and/or aerosol. Specificallycontemplated routes are oral administration, intravenous administration(e.g., systemic intravenous injection), regional administration viablood and/or lymph supply, and/or direct administration to an affectedsite. In general the most appropriate route of administration willdepend upon a variety of factors including the nature of the agent(e.g., its stability in the environment of the gastrointestinal tract),and/or the condition of the subject (e.g., whether the subject is ableto tolerate oral administration).

The exact amount of a compound required to achieve an effective amountwill vary from subject to subject, depending, for example, on species,age, and general condition of a subject, severity of the side effects ordisorder, identity of the particular compound(s), mode ofadministration, and the like. The desired dosage can be delivered threetimes a day, two times a day, once a day, every other day, every thirdday, every week, every two weeks, every three weeks, or every fourweeks. In certain embodiments, the desired dosage can be delivered usingmultiple administrations (e.g., two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, or moreadministrations).

In certain embodiments, an effective amount of a compound foradministration one or more times a day to a 70 kg adult human maycomprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg,about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosageform.

In certain embodiments, a compound described herein may be administeredat dosage levels sufficient to deliver from about 0.001 mg/kg to about1000 mg/kg, from about 0.01 mg/kg to about mg/kg, from about 0.1 mg/kgto about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, orfrom about 1 mg/kg to about 25 mg/kg, of subject body weight per day,one or more times a day, to obtain the desired therapeutic effect.

In some embodiments, a compound described herein is administered one ormore times per day, for multiple days. In some embodiments, the dosingregimen is continued for days, weeks, months, or years.

It will be appreciated that dose ranges as described herein provideguidance for the administration of provided pharmaceutical compositionsto an adult. The amount to be administered to, for example, a child oran adolescent can be determined by a medical practitioner or personskilled in the art and can be lower or the same as that administered toan adult.

It will be also appreciated that a compound or composition, as describedherein, can be administered in combination with one or more additionaltherapeutically active agents. In certain embodiments, a compound orcomposition provided herein is administered in combination with one ormore additional therapeutically active agents that improve itsbioavailability, reduce and/or modify its metabolism, inhibit itsexcretion, and/or modify its distribution within the body. It will alsobe appreciated that the therapy employed may achieve a desired effectfor the same disorder, and/or it may achieve different effects.

The compound or composition can be administered concurrently with, priorto, or subsequent to, one or more additional therapeutically activeagents. In certain embodiments, the additional therapeutically activeagent is a compound of Formula (A), e.g., Formula (I). In certainembodiments, the additional therapeutically active agent is not acompound of Formula (A), e.g., Formula (I). In general, each agent willbe administered at a dose and/or on a time schedule determined for thatagent. In will further be appreciated that the additionaltherapeutically active agent utilized in this combination can beadministered together in a single composition or administered separatelyin different compositions. The particular combination to employ in aregimen will take into account compatibility of a provided compound withthe additional therapeutically active agent and/or the desiredtherapeutic effect to be achieved. In general, it is expected thatadditional therapeutically active agents utilized in combination beutilized at levels that do not exceed the levels at which they areutilized individually. In some embodiments, the levels utilized incombination will be lower than those utilized individually.

Exemplary additional therapeutically active agents include, but are notlimited to, small organic molecules such as drug compounds (e.g.,compounds approved by the U.S. Food and Drug Administration as providedin the Code of Federal Regulations (CFR)), peptides, proteins,carbohydrates, monosaccharides, oligosaccharides, polysaccharides,nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides orproteins, small molecules linked to proteins, glycoproteins, steroids,nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides,antisense oligonucleotides, lipids, hormones, vitamins, and cells.

Also encompassed by the present disclosure are kits (e.g.,pharmaceutical packs). The kits provided may comprise a providedpharmaceutical composition or compound and a container (e.g., a vial,ampule, bottle, syringe, and/or dispenser package, or other suitablecontainer). In some embodiments, provided kits may optionally furtherinclude a second container comprising a pharmaceutical excipient fordilution or suspension of a provided pharmaceutical composition orcompound. In some embodiments, a provided pharmaceutical composition orcompound provided in the container and the second container are combinedto form one unit dosage form. In some embodiments, a provided kitsfurther includes instructions for use.

Compounds and compositions described herein are generally useful for theinhibition of PRMT5. In some embodiments, methods of treatingPRMT5-mediated disorder in a subject are provided which compriseadministering an effective amount of a compound described herein (e.g.,a compound of Formula (A), e.g., Formula (I)), or a pharmaceuticallyacceptable salt thereof), to a subject in need of treatment. In certainembodiments, the effective amount is a therapeutically effective amount.In certain embodiments, the effective amount is a prophylacticallyeffective amount. In certain embodiments, the subject is suffering froma PRMT5-mediated disorder. In certain embodiments, the subject issusceptible to a PRMT5-mediated disorder.

As used herein, the term “PRMT5-mediated disorder” means any disease,disorder, or other pathological condition in which PRMT5 is known toplay a role. Accordingly, in some embodiments, the present disclosurerelates to treating or lessening the severity of one or more diseases inwhich PRMT5 is known to play a role.

In some embodiments, the present disclosure provides a method ofinhibiting PRMT5 comprising contacting PRMT5 with an effective amount ofa compound described herein (e.g., a compound of Formula (A), e.g.,Formula (I)), or a pharmaceutically acceptable salt thereof. The PRMT5may be purified or crude, and may be present in a cell, tissue, orsubject. Thus, such methods encompass both inhibition of in vitro and invivo PRMT5 activity. In certain embodiments, the method is an in vitromethod, e.g., such as an assay method. It will be understood by one ofordinary skill in the art that inhibition of PRMT5 does not necessarilyrequire that all of the PRMT5 be occupied by an inhibitor at once.Exemplary levels of inhibition of PRMT5 include at least 10% inhibition,about 10% to about 25% inhibition, about 25% to about 50% inhibition,about 50% to about 75% inhibition, at least 50% inhibition, at least 75%inhibition, about 80% inhibition, about 90% inhibition, and greater than90% inhibition.

In some embodiments, provided is a method of inhibiting PRMT5 activityin a subject in need thereof comprising administering to the subject aneffective amount of a compound described herein (e.g., a compound ofFormula (A), e.g., Formula (I)), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof.

In certain embodiments, provided is a method of altering gene expressionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (A), e.g., Formula (I), or a pharmaceuticallyacceptable salt thereof. In certain embodiments, the cell in culture invitro. In certain embodiments, the cell is in an animal, e.g., a human.In certain embodiments, the cell is in a subject in need of treatment.

In certain embodiments, provided is a method of altering transcriptionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (A), e.g., Formula (I), or a pharmaceuticallyacceptable salt thereof. In certain embodiments, the cell in culture invitro. In certain embodiments, the cell is in an animal, e.g., a human.In certain embodiments, the cell is in a subject in need of treatment.

In certain embodiments, a method is provided of selecting a therapy fora subject having a disease associated with PRMT5-mediated disorder ormutation comprising the steps of determining the presence ofPRMT5-mediated disorder or gene mutation in the PRMT5 gene or andselecting, based on the presence of PRMT5-mediated disorder a genemutation in the PRMT5 gene a therapy that includes the administration ofa provided compound. In certain embodiments, the disease is cancer.

In certain embodiments, a method of treatment is provided for a subjectin need thereof comprising the steps of determining the presence ofPRMT5-mediated disorder or a gene mutation in the PRMT5 gene andtreating the subject in need thereof, based on the presence of aPRMT5-mediated disorder or gene mutation in the PRMT5 gene with atherapy that includes the administration of a provided compound. Incertain embodiments, the subject is a cancer patient.

In some embodiments, a provided compound is useful in treating aproliferative disorder, such as cancer, a benign neoplasm, an autoimmunedisease, or an inflammatory disease. For example, while not being boundto any particular mechanism, PRMT5 has been shown to be involved incyclin D1 dysregulated cancers. Increased PRMT5 activity mediates keyevents associated with cyclin D1-dependent neoplastic growth includingCUL4 repression, CDT1 overexpression, and DNA re-replication. Further,human cancers harboring mutations in Fbx4, the cyclin D1 E3 ligase,exhibit nuclear cyclin D1 accumulation and increased PRMT5 activity.See, e.g., Aggarwal et al., Cancer Cell. (2010) 18(4):329-40.Additionally, PRMT5 has also been implicated in accelerating cell cycleprogression through G1 phase and modulating regulators of G1; forexample, PRMT5 may upregulate cyclin-dependent kinase (CDK) 4, CDK6, andcyclins D1, D2 and E1. Moreover, PRMT5 may activate phosphoinositide3-kinase (PI3K)/AKT signaling. See, e.g., Wei et al., Cancer Sci. (2012)103(9):1640-50. PRMT5 has been reported to play a role in apoptosisthrough methylation of E2F-1. See, e.g., Cho et al., EMBO J. (2012)31:1785-1797; Zheng et al., Mol. Cell. (2013) 52:37-51. PRMT5 has beenreported to be an essential regulator of splicing and affect thealternative splicing of ‘sensor’ mRNAs that can then lead to defects indownstream events such as apoptosis. See, e.g., Bezzi et al., Genes Dev.(2013) 27:1903-1916. PRMT5 has been reported to play a role in theRAS-ERK pathway. See, e.g., Andrew-Perez et al., Sci Signal. (2011)September 13; 4(190)ra58 doi: 10.1126/scisignal.2001936. PRMT5 has beenreported to affect C/EBPb target genes through interaction with theMediator complex and hence affect cellular differentiation andinflammatory response. See, e.g., Tsutsui et al., J. Biol. Chem. (2013)288:20955-20965. PRMT5 has been shown to methylate HOXA9 essential forELAM expression during the EC inflammatory response. See, e.g.,Bandyopadhyay et al., Mol. Cell. Biol. (2012) 32:1202-1203. Thus in someembodiments, the inhibition of PRMT5 by a provided compound is useful intreating the following non-limiting list of cancers: breast cancer,esophageal cancer, bladder cancer, lung cancer, hematopoietic cancer,lymphoma, medulloblastoma, rectum adenocarcinoma, colon adenocarcinoma,gastric cancer, pancreatic cancer, liver cancer, adenoid cysticcarcinoma, lung adenocarcinoma, head and neck squamous cell carcinoma,brain tumors, hepatocellular carcinoma, renal cell carcinoma, melanoma,oligodendroglioma, ovarian clear cell carcinoma, and ovarian serouscystadenocarcinoma. See, e.g., Pal et al., EMBO J. (2007) 26:3558-3569(mantle cell lymphoma); Wang et al., Mol. Cell Biol. (2008) 28:6262-77(chronic lymphocytic leukemia (CLL)); and Tae et al., Nucleic Acids Res.(2011) 39:5424-5438.

In some embodiments, the inhibition of PRMT5 by a provided compound isuseful in treating prostate cancer and lung cancer, in which PRMT5 hasbeen shown to play a role. See, e.g., Gu et al., PLoS One 2012;7(8):e44033; Gu et al., Biochem. J. (2012) 446:235-241. In someembodiments, a provided compound is useful to delay the onset of, slowthe progression of, or ameliorate the symptoms of cancer. In someembodiments, a provided compound is administered in combination withother compounds, drugs, or therapeutics to treat cancer.

In some embodiments, compounds described herein are useful for treatinga cancer including, but not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma),appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g.,cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast), brain cancer (e.g., meningioma;glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchuscancer, carcinoid tumor, cervical cancer (e.g., cervicaladenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma,colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma(e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma),endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophagealcancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),Ewing sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma),familiar hypereosinophilia, gall bladder cancer, gastric cancer (e.g.,stomach adenocarcinoma), gastrointestinal stromal tumor (GIST), head andneck cancer (e.g., head and neck squamous cell carcinoma, oral cancer(e.g., oral squamous cell carcinoma (OSCC), throat cancer (e.g.,laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer,oropharyngeal cancer)), hematopoietic cancers (e.g., leukemia such asacute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acutemyelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronicmyelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chroniclymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma suchas Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkinlymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma(DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicularlymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas(e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma),primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacyticlymphoma (i.e., “Waldenström's macroglobulinemia”), hairy cell leukemia(HCL), immunoblastic large cell lymphoma, precursor B-lymphoblasticlymphoma and primary central nervous system (CNS) lymphoma; and T-cellNHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheralT-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma,extranodal natural killer T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplasticlarge cell lymphoma); a mixture of one or more leukemia/lymphoma asdescribed above; and multiple myeloma (MM)), heavy chain disease (e.g.,alpha chain disease, gamma chain disease, mu chain disease),hemangioblastoma, inflammatory myofibroblastic tumors, immunocyticamyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor,renal cell carcinoma), liver cancer (e.g., hepatocellular cancer (HCC),malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, smallcell lung cancer (SCLC), non-small cell lung cancer (NSCLC),adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g.,systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma,myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV),essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocyticleukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilicsyndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis(NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g.,gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoidtumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarianembryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma,pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductalpapillary mucinous neoplasm (IPMN), Islet cell tumors), penile cancer(e.g., Paget's disease of the penis and scrotum), pinealoma, primitiveneuroectodermal tumor (PNT), prostate cancer (e.g., prostateadenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer,skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA),melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g.,appendix cancer), soft tissue sarcoma (e.g., malignant fibroushistiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor(MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous glandcarcinoma, sweat gland carcinoma, synovioma, testicular cancer (e.g.,seminoma, testicular embryonal carcinoma), thyroid cancer (e.g.,papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC),medullary thyroid cancer), urethral cancer, vaginal cancer, and vulvarcancer (e.g., Paget's disease of the vulva).

In some embodiments, a provided compound is useful in treating ametabolic disorder, such as diabetes or obesity. For example, while notbeing bound to any particular mechanism, a role for PRMT5 has beenrecognized in adipogenesis. Inhibition of PRMT5 expression in multiplecell culture models for adipogenesis prevented the activation ofadipogenic genes, while overexpression of PRMT5 enhanced adipogenic geneexpression and differentiation. See, e.g., LeBlanc et al., MolEndocrinol. (2012) 26:583-597. Additionally, it has been shown thatadipogenesis plays a pivotal role in the etiology and progression ofdiabetes and obesity. See, e.g., Camp et al., Trends Mol Med. (2002)8:442-447. Thus in some embodiments, the inhibition of PRMT5 by aprovided compound is useful in treating diabetes and/or obesity.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, diabetes. Insome embodiments, the diabetes is Type 1 diabetes. In some embodiments,the diabetes is Type 2 diabetes. In some embodiments, a providedcompound is useful to delay the onset of, slow the progression of, orameliorate the symptoms of, obesity. In some embodiments, a providedcompound is useful to help a subject lose weight. In some embodiments, aprovided compound could be used in combination with other compounds,drugs, or therapeutics, such as metformin and insulin, to treat diabetesand/or obesity.

In some embodiments, a provided compound is useful in treating a blooddisorder, e.g., a hemoglobinopathy, such as sickle cell disease orβ-thalassemia. For example, while not being bound to any particularmechanism, PRMT5 is a known repressor of γ-globin gene expression, andincreased fetal γ-globin (HbF) levels in adulthood are associated withsymptomatic amelioration in sickle cell disease and β-thalassemia. See,e.g., Xu et al., Haematologica. (2012) 97:1632-1640; Rank et al. Blood.(2010) 116:1585-1592. Thus in some embodiments, the inhibition of PRMT5by a provided compound is useful in treating a blood disorder, such as ahemoglobinopathy such as sickle cell disease or β-thalassemia.

In some embodiments, a provided compound is useful to delay the onsetof, slow the progression of, or ameliorate the symptoms of, sickle celldisease. In some embodiments, a provided compound is useful to delay theonset of, slow the progression of, or ameliorate the symptoms of,β-thalassemia. In some embodiments, a provided compound could be used incombination with other compounds, drugs, or therapeutics, to treat ahemoglobinopathy such as sickle cell disease or β-thalassemia.

In some embodiments, a provided compound is useful in treatinginflammatory and autoimmune disease. PRMT5 is reported to activate NFkBsignaling pathway through the methylation of p65. PRMT5 is reported tointeract with Death receptor 4 and Death receptor 5 contributing toTRAIL-induced activation of inhibitor or kB kinase (IKK) and nuclearfactor-kB (NF-kB). See, e.g., Tanaka et al., Mol. Cancer. Res. (2009)7:557-569; Wei et al., Proc. Nat'l. Acad. Sci. USA (2013) 110:13516-21.

The term “inflammatory disease” refers to those diseases, disorders orconditions that are characterized by signs of pain (dolor, from thegeneration of noxious substances and the stimulation of nerves), heat(calor, from vasodilatation), redness (rubor, from vasodilatation andincreased blood flow), swelling (tumor, from excessive inflow orrestricted outflow of fluid), and/or loss of function (functio laesa,which can be partial or complete, temporary or permanent. Inflammationtakes on many forms and includes, but is not limited to, acute,adhesive, atrophic, catarrhal, chronic, cirrhotic, diffuse,disseminated, exudative, fibrinous, fibrosing, focal, granulomatous,hyperplastic, hypertrophic, interstitial, metastatic, necrotic,obliterative, parenchymatous, plastic, productive, proliferous,pseudomembranous, purulent, sclerosing, seroplastic, serous, simple,specific, subacute, suppurative, toxic, traumatic, and/or ulcerativeinflammation.

Exemplary inflammatory diseases include, but are not limited to,inflammation associated with acne, anemia (e.g., aplastic anemia,haemolytic autoimmune anaemia), asthma, arteritis (e.g., polyarteritis,temporal arteritis, periarteritis nodosa, Takayasu's arteritis),arthritis (e.g., crystalline arthritis, osteoarthritis, psoriaticarthritis, gouty arthritis, reactive arthritis, rheumatoid arthritis andReiter's arthritis), ankylosing spondylitis, amylosis, amyotrophiclateral sclerosis, autoimmune diseases, allergies or allergic reactions,atherosclerosis, bronchitis, bursitis, chronic prostatitis,conjunctivitis, Chagas disease, chronic obstructive pulmonary disease,cermatomyositis, diverticulitis, diabetes (e.g., type I diabetesmellitus, type 2 diabetes mellitus), a skin condition (e.g., psoriasis,eczema, burns, dermatitis, pruritus (itch)), endometriosis,Guillain-Barre syndrome, infection, ischaemic heart disease, Kawasakidisease, glomerulonephritis, gingivitis, hypersensitivity, headaches(e.g., migraine headaches, tension headaches), ileus (e.g.,postoperative ileus and ileus during sepsis), idiopathicthrombocytopenic purpura, interstitial cystitis (painful bladdersyndrome), gastrointestinal disorder (e.g., selected from peptic ulcers,regional enteritis, diverticulitis, gastrointestinal bleeding,eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis,eosinophilic gastritis, eosinophilic gastroenteritis, eosinophiliccolitis), gastritis, diarrhea, gastroesophageal reflux disease (GORD, orits synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn'sdisease, ulcerative colitis, collagenous colitis, lymphocytic colitis,ischaemic colitis, diversion colitis, Behcet's syndrome, indeterminatecolitis) and inflammatory bowel syndrome (IBS)), lupus, multiplesclerosis, morphea, myeasthenia gravis, myocardial ischemia, nephroticsyndrome, pemphigus vulgaris, pernicious aneaemia, peptic ulcers,polymyositis, primary biliary cirrhosis, neuroinflammation associatedwith brain disorders (e.g., Parkinson's disease, Huntington's disease,and Alzheimer's disease), prostatitis, chronic inflammation associatedwith cranial radiation injury, pelvic inflammatory disease, reperfusioninjury, regional enteritis, rheumatic fever, systemic lupuserythematosus, schleroderma, scierodoma, sarcoidosis,spondyloarthopathies, Sjogren's syndrome, thyroiditis, transplantationrejection, tendonitis, trauma or injury (e.g., frostbite, chemicalirritants, toxins, scarring, burns, physical injury), vasculitis,vitiligo and Wegener's granulomatosis.

In certain embodiments, the inflammatory disease is an acuteinflammatory disease (e.g., for example, inflammation resulting frominfection). In certain embodiments, the inflammatory disease is achronic inflammatory disease (e.g., conditions resulting from asthma,arthritis and inflammatory bowel disease). The compounds may also beuseful in treating inflammation associated with trauma andnon-inflammatory myalgia. The compounds may also be useful in treatinginflammation associated with cancer.

Exemplary autoimmune diseases, include, but are not limited to,arthritis (including rheumatoid arthritis, spondyloarthopathies, goutyarthritis, degenerative joint diseases such as osteoarthritis, systemiclupus erythematosus, Sjogren's syndrome, ankylosing spondylitis,undifferentiated spondylitis, Behcet's disease, haemolytic autoimmuneanaemias, multiple sclerosis, amyotrophic lateral sclerosis, amylosis,acute painful shoulder, psoriatic, and juvenile arthritis), asthma,atherosclerosis, osteoporosis, bronchitis, tendonitis, bursitis, skincondition (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itch)),enuresis, eosinophilic disease, gastrointestinal disorder (e.g.,selected from peptic ulcers, regional enteritis, diverticulitis,gastrointestinal bleeding, eosinophilic gastrointestinal disorders(e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilicgastroenteritis, eosinophilic colitis), gastritis, diarrhea,gastroesophageal reflux disease (GORD, or its synonym GERD),inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerativecolitis, collagenous colitis, lymphocytic colitis, ischaemic colitis,diversion colitis, Behcet's syndrome, indeterminate colitis) andinflammatory bowel syndrome (IBS)), and disorders ameliorated by agastroprokinetic agent (e.g., ileus, postoperative ileus and ileusduring sepsis; gastroesophageal reflux disease (GORD, or its synonymGERD); eosinophilic esophagitis, gastroparesis such as diabeticgastroparesis; food intolerances and food allergies and other functionalbowel disorders, such as non-ulcerative dyspepsia (NUD) and non-cardiacchest pain (NCCP, including costo-chondritis)).

In some embodiments, a provided compound is useful in somatic cellreprogramming, such as reprogramming somatic cells into stem cells. See,e.g., Nagamatsu et al., J Biol Chem. (2011) 286:10641-10648. In someembodiments, a provided compound is useful in germ cell development, andare thus envisioned useful in the areas of reproductive technology andregenerative medicine. See, e.g., Ancelin et al., Nat. Cell. Biol.(2006) 8:623-630.

In some embodiments, compounds described herein can prepared usingmethods shown in general Scheme 1, which describes ring opening of achiral or racemic epoxide group to form the amino alcohol moiety linker.Further substitution of the tetrahydroisoquinoline ring and/or theG-containing ring can be carried out before or after the couplingreaction.

In some embodiments, epoxide opening can be performed in the final stepas shown in exemplary Schemes 2 and 3.

In some embodiments, an amide coupling step can be used to provide a keyintermediate for further synthesis, as shown in exemplary Schemes 4-6.

In some embodiments of the compounds described herein, R¹² or R¹³ is anamine. A non-limiting example of the synthetic sequence used to preparesuch analogs is provided herein (see Scheme 7). In this example, analcohol of Formula (Z-1) is oxidized under suitable conditions S1 toaffect transformation into an intermediate ketone of Formula (Z-2). Aketone of Formula (Z-2) can be contacted with a primary or secondaryamine under suitable conditions S2 to affect a reductive amination whichcan afford an amino compound of Formula (Z-3).

In some embodiments, the oxidation reaction S1 is carried out directlywith a stoichiometeric oxidant. In some embodiments, the stoichiometricoxidant is pyridinium chlorochromate. In some embodiments, thestoichiometric oxidant is pyridinium dichromate. In some embodiments,the stoichiometric oxidant is Dess-Martin periodinane. In someembodiments, the stoichiometric oxidant is prepared in situ. In someembodiments, the stoichiometric oxidant is prepared in situ using sulfurtrioxide pyridine complex and dimethylsulfoxide. In some embodiments,the stoichiometric oxidant is prepared in situ using oxallyl chlorideand dimethylsulfoxide. In some embodiments, the stoichiometric oxidantis prepared in situ using a carbodiimide and dimethylsulfoxide. In someembodiments, the stoichiometric oxidant is prepared in situ usingN-chlorosuccinimide and dimethylsulfide. In some embodiments, theoxidation reaction S1 is catalyzed. In some embodiments, the catalyst is(2,2,6,6-tetramethyl-piperidin-1-yl)oxyl. In some embodiments, thecatalyst is a ruthenium complex. In some embodiments, the catalyst is apalladium complex. In some embodiments, the catalyst is a coppercomplex. For examples of standard methods and conditions for alcoholoxidation, see Epstein et al., Chem. Rev. (1967) 67(3):247-260 and B. M.Trost ed. “Comprehensive Organic Synthesis”, (1991), Vol. 7, p 281-305.

In some embodiments, both the oxidation step S1 and reductive aminationstep S2 occur in one pot. In some embodiments, both the oxidation stepS1 and the reductive amination step S2 are carried out using the samecatalyst. In some embodiments, the catalyst is a rhodium complex. Insome embodiments, the catalyst is a ruthenium complex. In someembodiments, the catalyst is an iridium complex.

In some embodiments, the reductive amination reaction S2 is carried outusing a borohydride. In some embodiments, the reductive aminationreaction S2 is carried out using sodium borohydride. In someembodiments, the reductive amination reaction S2 is carried out usingsodium cyanoborohydride. In some embodiments, the reductive aminationreaction S2 is carried out using sodium triacetoxyborohydride. In someembodiments, the reductive amination reaction S2 is carried out using aborane. In some embodiments, the reductive amination reaction S2 iscarried out using a silyl hydride. In some embodiments, the reductiveamination reaction S2 is carried out using hydrogen. In someembodiments, the reductive amination reaction S2 is carried out in twosteps, by first contacting a ketone of (Z-2) with an amine to form anintermediate imine, and then reducing the intermediate imine undersufficient conditions to afford a compound of Formula (Z-3). In someembodiments, the reaction conditions S2 comprise addition of a proticacid. In some embodiments, the reaction conditions S2 comprise additionof an aprotic acid. In some embodiments, the reaction conditions S2comprise in situ formation of the reducing agent. In some embodiments,the reaction conditions S2 comprise a catalyst. In some embodiments, thereaction conditions S2 comprise a transition metal catalyst. In someembodiments, the reaction conditions S2 comprise a palladium or nickelcatalyst. In some embodiments, the reductive amination reaction S2 isstereoselective. In some embodiments, the stereoselective reductiveamination reaction S2 is carried out in the presence of a chiralcatalyst. For examples of standard methods and conditions for reductiveaminations, see Gomez et al., Adv. Synth. Catal. (2002)344(10):1037-1057 and Abdel-Magid et al., J. Org. Chem. (1996), 61:3849.

An alternative non-limiting synthetic sequence leading to theaforementioned amine analogs is described herein (see Scheme 8). Thehydroxyl moiety of a compound of Formula (Z-4) can be transformed into aleaving group under sufficient conditions S3 to afford a compound ofFormula (Z-5). The leaving group of a compound of Formula (Z-5) can bedisplaced with an amine under suitable conditions S4 to produce an aminocompound of Formula (Z-6).

In some embodiments, LG of Formula (Z-5) is a halide. In someembodiments, LG of Formula (Z-5) is bromine. In some embodiments, LG ofFormula (Z-5) is iodine. In some embodiments, LG of Formula (Z-5) is asubstituted or unsubstituted alkyl sulfonate. In some embodiments, LG ofFormula (Z-5) is a substituted or unsubstituted aryl sulfonate. In someembodiments, LG of Formula (Z-5) is methyl sulfonate. In someembodiments, LG of Formula (A-5) is trifluoromethane sulfonate. In someembodiments, LG of Formula (Z-5) is a toluene sulfonate. In someembodiments, LG of Formula (Z-5) is a nitrobenzene sulfonate. In someembodiments, when LG of Formula (Z-5) is halide, conditions S3 comprisea phosphoryl halide. In some embodiments, when LG of Formula (Z-5) ishalide, conditions S3 comprise a sulfuryl halide. In some embodiments,when LG of Formula (Z-5) is sulfonate, conditions S3 comprise a sulfonylhalide. In some embodiments, when LG of Formula (Z-5) is sulfonate,conditions S3 comprise a sulfonyl anhydride. For examples of standardmethods and conditions for organohalide or sulfonate ester synthesis,see Lautens et al., Synthesis (2011) 2:342-346 or Marcotullio et al.,Synthesis (2006) 16:2760-2766.

In some embodiments, conditions S4 are neutral. In some embodiments,conditions S4 comprise addition of a base. In certain embodiments ofconditions S4, the base is either inorganic or organic. In certainembodiments of conditions S4, the base is inorganic. In certainembodiments of conditions S4, the base is organic. In certainembodiments of conditions S4, the base is a metal acetate, alkoxide,amide, amidine, carbonate, hydroxide, phenoxide, or phosphate. Incertain embodiments of conditions S4, the base is sodium, potassium, orcaesium carbonate. In certain embodiments of conditions S4, the base issodium, potassium, or caesium bicarbonate. In certain embodiments ofconditions S4, the base is 1,1,3,3-tetramethylguanidine,1,4-diazabicyclo[2.2.2]octane, 1,8-bis(dimethylamino)naphthalene,1,8-diazabicycloundec-7-ene, ammonia, diisopropylamine, imidazole,N,N-diisopropylethylamine, piperidine, pyridine, pyrrolidine, ortriethylamine. In some embodiments of conditions S4, the solvent is apolar protic solvent. In some embodiments of conditions S4, the solventis a polar aprotic solvent. In some embodiments of conditions S4, thereaction is performed in the absence of solvent. In some embodiments,conditions S4 comprise a catalyst. In some embodiments of conditions S4,the catalyst is an iodide salt. In some embodiments, both step S3 andthe displacement step S4 occur in one pot. In some embodiments, thehydroxyl moiety of a compound of Formula (Z-4) is converted into aleaving group in situ. In some embodiments, the hydroxyl moiety of acompound of Formula (Z-4) is converted into a leaving group in situusing an azodicarboxylate and an aryl or alkyl phosphine. For examplesof standard methods and conditions for amine syntheses throughalkylation reactions, see Salvatore et. al, Tetrahedron (2001)57:7785-7811.

EXAMPLES

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Synthetic Methods Intermediate Synthesis2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 1,2,3,4-tetrahydroisoquinoline (15 g, 0.11 mol) in MeCN(100 mL) was added K₂CO₃ (30.7 g, 0.23 mol) at 0° C.2-(bromomethyl)oxirane (17 g, 0.12 mol) was added to the reaction after1 h. The solution was stirred at 22° C. for 16 h at which time thesolids were filtered and washed with MeCN. The solution was concentratedand the residue was used in the next step without further purification(17 g, Yield 78%). LCMS (m/z): 190.1 (M+1).

Compound 11-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)indolin-2-one

To a solution of indolin-2-one (50.0 mg, 0.378 mmol) in ethanol (5 mL)was added 2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline (85.0 mg,0.451 mmol). The mixture was heated under microwave conditions at 120°C. for 0.5 h, concentrated and purified by preparative HPLCpurification. (40.0 mg, yield 33.0%) MS (ESI⁺) e/z: 323.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 7.30-7.23 (m, 2H), 7.14-7.07 (m, 4H), 7.06 (d,J=7.5 Hz, 1H), 7.01 (t, J=5.8 Hz, 1H), 4.94 (d, J=7.8 Hz, 2H), 4.31-4.18(m, 1H), 3.97-3.86 (m, 1H), 3.82-3.72 (m, 1H), 3.71-3.64 (m, 2H),2.90-2.73 (m, 4H), 2.73-2.59 (m, 2H).

Compound 31-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(m-tolyl)piperazin-2-one

Step 1: tert-butyl 3-oxopiperazine-1-carboxylate

Piperazin-2-one (10 g, 0.1 mol), Et₃N (20.2 g, 0.2 mmol) and DCM (100mL) were combined and cooled to 0° C. To the solution was added Boc₂O(26.1 g, 0.12 mol) in DCM (250 ml), and the resulting mixture was warmedto room temperature and stirred for 12 h. The reaction mixture wasconcentrated and dissolved in ethyl acetate, washed with 1 N HCl, brine,dried over sodium sulfate, filtered and concentrated. The crude productwas used without further purification. (19.5 g, yield 97.5%).

Step 2: tert-butyl 4-(oxiran-2-ylmethyl)-3-oxopiperazine-1-carboxylate

tert-butyl 3-oxopiperazine-1-carboxylate (400 mg, 2 mmol) was dissolvedin THF (20 mL) and cooled to 0° C. NaH (72 mg, 3 mmol) was added to thesolution followed by 2-(bromomethyl)oxirane (326 mg, 2.4 mmol), and theresulting mixture was stirred at 0° C. for 16 h. The reaction mixturewas quenched by addition of ice water, extracted with ethyl acetate,washed with brine, dried over sodium sulfate and concentrated. The crudeproduct was used without further purification. (400 mg, yield 78.1%).

Step 3: tert-butyl4-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-3-oxopiperazine-1-carboxylate

To a solution of tert-butyl4-(oxiran-2-ylmethyl)-3-oxopiperazine-1-carboxylate (400 mg, 1.56 mmol)in EtOH (1 mL) was added 1,2,3,4-tetrahydroisoquinoline (207 mg, 1.56mmol). The mixture was heated under microwave conditions at 120° C. for0.5 h and concentrated. The crude product was used without furtherpurification. (600 mg, yield 98%) MS (ESI⁺) e/z: 390.2 [M+1]⁺.

Step4:1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one

To a solution of tert-butyl4-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxylpropyl)-3-oxopiperazine-1-carboxylate (200 mg, 0.69 mmol) in ethylacetate (10 mL) was added HCl/ethyl acetate (4 mL). The reaction mixturewas stirred at 20° C. overnight, concentrated and used in the next stepwithout further purification. (100 mg, yield 50%) MS (ESI⁺) e/z: 290.2[M+1]⁺.

Step5:1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(m-tolyl)piperazin-2-one

A mixture of1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one(50 mg, 0.17 mmol), 1-iodo-3-methylbenzene (55.3 mg, 0.25 mmol),Pd₂(dba)₃ (10 mg, 0.01 mmol), Xantphos (20 mg, 0.02 mmol) and t-BuONa(30 mg, 0.3 mmol) were placed in the three-neck reactor with a condenserand magnetic stirring bar, the system was degassed 3 times (N₂) anddioxane (10 mL) was added. The mixture was heated at reflux for 16 h,cooled, filtered and concentrated. The residue was purified bypreparative HPLC purification. (18.0 mg, yield 28.0%) MS (ESI⁺) e/z:380.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.10 (br. s., 5H),6.85-6.65 (m, 3H), 4.33-4.14 (m, 1H), 3.88-3.81 (m, 2H), 3.81-3.68 (m,4H), 3.67-3.58 (m, 1H), 3.56-3.44 (m, 2H), 3.32-3.25 (m, 1H), 3.02-2.76(m, 4H), 2.68-2.54 (m, 2H), 2.32 (s, 3H).

Compound 41-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(quinolin-8-yl)piperazin-2-one

To a solution of 1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one (100 mg, 0.35 mmol) in 10 mL of dry toluene was added8-bromoquinoline (108 mg, 0.52 mmol), Pd₂(dba)₃ (32 mg, 0.035 mmol),NINAP (44 mg, 0.07 mmol) and t-BuONa (101 mg, 1.05 mmol). The flask wasdegassed and charged with N₂, then heated at 110° C. for 16 h. Thereaction mixture was cooled, filtered, and concentrated. The residue waspurified by preparative HPLC purification. (23 mg, yield 15.8%) MS(ESI⁺) e/z: 417.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 8.96-8.85 (m,1H), 8.53-8.39 (m, 1H), 8.38-8.29 (m, 1H), 7.70-7.62 (m, 1H), 7.62-7.51(m, 2H), 7.37-7.24 (m, 4H), 7.24-7.17 (m, 1H), 4.60-4.52 (m, 1H),4.52-4.41 (m, 2H), 4.08-3.98 (m, 2H), 3.87-3.78 (m, 2H), 3.77-3.69 (m,1H), 3.69-3.58 (m, 4H), 3.57-3.49 (m, 1H), 3.42-3.34 (m, 1H), 3.32-3.27(m, 1H), 3.27-3.13 (m, 2H).

Compound 54-(3-chlorophenyl)-1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one

Compound 5 was synthesized in by a procedure analogous to that employedfor compound 3. (5.3 mg, yield 7.8%) MS (ESI⁺) e/z: 400.1 [M+1]⁺. ¹H NMR(MeOD, 400 MHz), δ ppm: 7.26-7.21 (m, 1H), 7.16-7.07 (m, 3H), 7.07-7.01(m, 1H), 6.95-6.89 (m, 1H), 6.89-6.78 (m, 2H), 4.26-4.19 (m, 1H),3.92-3.85 (m, 2H), 3.83-3.70 (m, 4H), 3.69-3.62 (m, 1H), 3.59-3.51 (m,2H), 3.32-3.24 (m, 1H), 2.97-2.90 (m, 2H), 2.89-2.80 (m, 2H), 2.66-2.56(m, 2H).

Compound 74-(4-chlorophenyl)-1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one

Compound 7 was synthesized in by a procedure analogous to that employedfor compound 3. (14.6 mg, yield 21.5%) MS (ESI⁺) e/z: 400.1[M+1]⁺. ¹HNMR (MeOD, 400 MHz), δ ppm: 7.28-7.16 (m, 2H), 7.16-6.98 (m, 4H),6.95-6.85 (m, 2H), 4.25-4.16 (m, 1H), 3.90-3.81 (m, 2H), 3.81-3.68 (m,4H), 3.67-3.58 (m, 1H), 3.55-3.45 (m, 2H), 2.97-2.87 (m, 2H), 2.86-2.75(m, 2H), 2.66-2.52 (m, 2H).

Compound 81-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(5-morpholinopyridin-3-yl)piperazin-2-one

Step 1: 4-(5-bromopyridin-3-yl)morpholine

To a solution of morpholine (3.67 g, 4.23 mmol), NMP (2 mL) and toluene(4 mL) was added 3,5-dibromopyridine (1.00 g, 4.20 mmol) and the mixturewas heated under microwave conditions at 180° C. for 1 h. The solutionwas partitioned between DCM and water, the organic layer was washed withwater, brine, dried over anhydrous sodium sulfate, filtered andconcentrated. The residue was purified by column chromatography elutingwith (EtOAc: PE=1:5). (200 mg, yield 19.5%) MS (ESI⁺) e/z: 243.0 [M+1]⁺LCMS.

Step 2:1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(5-morpholinopyridin-3-yl)piperazin-2-one

A solution of 4-(5-bromopyridin-3-yl)morpholine (100 mg, 0.413 mmol),1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one(119 mg, 0.413 mmol), X-phos (20 mg, 0.042 mmol), Pd(dba)₂ (10 mg, 0.017mmol) and NaOtBu (59.0 mg, 0.615 mmol) in dioxane (5 mL) was heated at120° C. 1 h under microwave conditions. The crude reaction mixture wasfiltered, concentrated, and purified by preparative HPLC purification.(6.10 mg, yield 3.3%) 1H NMR (MeOD, 400 MHz), δ ppm: 7.76 (d, J=2.3 Hz,1H), 7.72 (d, J=2.0 Hz, 1H), 7.17-7.07 (m, 3H), 7.07-7.01 (m, 1H),6.89-6.80 (m, 1H), 4.28-4.18 (m, 1H), 3.93 (s, 2H), 3.89-3.82 (m, 4H),3.82-3.72 (m, 4H), 3.71-3.63 (m, 1H), 3.59 (t, J=1.0 Hz, 2H), 3.32-3.28(m, 1H), 3.27-3.16 (m, 4H), 2.99-2.90 (m, 2H), 2.90-2.81 (m, 2H),2.68-2.58 (m, 2H).

Compound 111-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(2-fluorophenyl)piperazin-2-one

Compound 11 was synthesized in by a procedure analogous to that employedfor compound 3. (15.7 mg, yield 24.1%) MS (ESI⁺) e/z: 384.2[M+1]⁺. ¹HNMR (MeOD, 400 MHz), δ ppm: 7.14-6.98 (m, 8H), 4.27-4.18 (m, 1H),3.81-3.68 (m, 6H), 3.65-3.57 (m, 1H), 3.43-3.36 (m, 2H), 3.30-3.22 (m,1H), 2.95-2.88 (m, 2H), 2.87-2.80 (m, 2H), 2.64-2.53 (m, 2H).

Compound 121-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(3-morpholinophenyl)piperazin-2-one

Step 1: 4-(3-bromophenyl)morpholine

To a solution of 3-bromoaniline (1.00 g, 5.81 mmol) in anhydrous DMF (20mL) was added 1-bromo-2-(2-bromoethoxyl)ethane (1.62 g, 6.18 mmol) andDIPEA (2.25 g, 17.4 mmol). The solution was heated to 100° C. for 16 h,cooled and concentrated. The residue was purified by preparative HPLCpurification. (230 mg, yield 16.4%) MS (ESI⁺) e/z: 242.0 [M+1]⁺.

Step 2:1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(3-morpholinophenyl)piperazin-2-one

4-(3-bromophenyl)morpholine (100 mg, 0.413 mmol),1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)piperazin-2-one(119 mg, 0.413 mmol), Xphos (20 mg, 0.042 mmol), Pd(dba)2 (10 mg, 0.017mmol) and NaOtBu (59.0 mg, 0.615 mmol) and dioxane (5 mL) were combinedin a three-neck flask with a condenser and magnetic stirring bar, thesystem was degassed (N₂) and dioxane (10 mL) was added. The reactionmixture was heated at 100° C. for 16 h, filtered and concentrated. Theresidue was purified by preparative HPLC purification. (3.2 mg, yield1.7%). ¹H NMR (MeOD, 400 MHz), δ ppm: 7.21-7.15 (m, 1H), 7.15-7.07 (m,3H), 7.07-7.00 (m, 1H), 6.58-6.45 (m, 3H), 4.22 (br. s., 1H), 3.86-3.82(m, 6H), 3.78-3.69 (m, 4H), 3.68-3.59 (m, 1H), 3.56-3.46 (m, 2H),3.31-3.26 (m, 1H), 3.16-3.12 (m, 4H), 2.98-2.90 (m, 2H), 2.90-2.78 (m,2H), 2.60 (d, J=6.0 Hz, 2H).

Compound 151-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(p-tolyl)piperazin-2-one

Compound 15 was synthesized in by a procedure analogous to that employedfor compound 3. (14.9 mg, yield 23.1%) MS (ESI⁺) e/z: 380.2 [M+1]⁺. ¹HNMR (MeOD, 400 MHz), δ ppm: 7.14-6.98 (m, 6H), 6.89-6.80 (m, 2H),4.26-4.13 (m, 1H), 3.84-3.65 (m, 6H), 3.65-3.55 (m, 1H), 3.49-3.38 (m,2H),) 3.30-3.20 (m, 1H), 2.96-2.86 (m, 2H), 2.86-2.76 (m, 2H), 2.64-2.53(m, 2H), 2.24 (s, 3H).

Compound 19(R)-1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(phenylamino)pyridin-2(1H)-one

Step 1: (R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline

A solution of 1,2,3,4-tetrahydroisoquinoline (10 g, 0.15 mol) in THF(100 mL) was cooled to 0° C. and potassium fluoride (22 g, 0.3 mol) wasadded. After 1 h, (R)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (21.4 g,0.17 mol) was added; the mixture was warmed to 25° C. and stirred for 16h. The solution was filtered, washed with THF and concentrated. Thecrude product was used in next step without further purification. (15 g,yield 53%) MS (ESI⁺) e/z: 190.1 [M+1]⁺.

Step 2:(R)-4-bromo-1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyridin-2(1H)-one

To a solution of 4-bromopyridin-2(1H)-one (1 g, 5.747 mmol) in EtOH (3mL) was added (R)-2-(oxiran-2-ylmethyl)-1,2,3,4-tetrahydroisoquinoline(1.087 g, 5.747 mmol). The mixture was heated under microwave conditionsat 120° C. for 0.5 h and concentrated. The crude product was purified bycolumn chromatography eluting with DCM/MeOH (10:1). (475 mg, yield22.8%) MS (ESI⁺) e/z: 363.1 [M+1]⁺.

Step 3:(R)-1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-4-(phenylamino)pyridin-2(1H)-one

To a solution of(R)-4-bromo-1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyridin-2(1H)-one(200 mg, 0.551 mmol) in anhydrous toluene (5 mL) was added aniline (51mg, 0.551 mmol), xantphos (20 mg), Pd₂(dba)₃ (20 mg) and t-BuOK (123 mg,1.102 mmol). The mixture was heated under microwave conditions at 140°C. for 1 h, filtered and concentrated. The crude product was purified bypreparative HPLC purification. (18 mg, yield 8.7%) MS (ESI⁺) e/z: 376.1[M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.45-7.35 (m, 3H), 7.24-7.18 (m,2H), 7.17-7.08 (m, 4H), 7.07-7.02 (m, 1H), 6.11 (dd, J=2.6, 7.5 Hz, 1H),5.99 (d, J=2.5 Hz, 1H), 4.33 (dd, J=3.3, 13.4 Hz, 1H), 4.27-4.15 (m,1H), 3.73 (s, 2H), 3.63 (dd, J=8.0, 13.6 Hz, 1H), 2.96-2.89 (m, 2H),2.88-2.82 (m, 2H), 2.66-2.58 (m, 2H).

Compound 314-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2H-benzo[b][1,4]oxazin-3(4H)-one

Step 1: ethyl 2-(2-iodophenoxyl)acetate

To a solution of 2-iodophenol (1 g, 4.55 mmol) in CH₃CN (20 mL) wasadded K₂CO₃ (0.93 g, 6.83 mmol) and ethyl 2-bromoacetate (0.835 g, 5.0mmol). The reaction mixture was stirred at reflux for 2 h, cooled toroom temperature, extracted with EtOAc, washed with brine, dried oversodium sulfate and concentrated. The crude product was used in the nextstep without further purification. (1.1 g, yield 79%) MS (ESI⁺) e/z:307.1 [M+1]⁺.

Step 2:N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-iodophenoxyl)acetamide

To a solution of ethyl 2-(2-iodophenoxyl)acetate (900 mg, 2.94 mmol) inEtOH (3 mL) was added1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (677 mg, 3.24mmol). The reaction mixture was heated under microwave conditions at 80°C. for 3 h, cooled and concentrated. The crude product was used in thenext step without further purification. (1.3 g, yield 95%) MS (ESI⁺)e/z: 467.1 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.80 (dd, J=1.0, 7.8Hz, 1H), 7.41-7.30 (m, 1H), 7.15-7.05 (m, 3H), 7.05-6.99 (m, 1H), 6.96(d, J=8.0 Hz, 1H), 6.87-6.76 (m, 1H), 4.66-4.53 (m, 2H), 4.06 (quin,J=5.9 Hz, 1H), 3.77-3.63 (m, 2H), 3.58 (dd, J=4.5, 13.6 Hz, 1H), 3.38(dd, J=6.4, 13.7 Hz, 1H), 2.98-2.86 (m, 2H), 2.86-2.76 (m, 2H),2.66-2.57 (m, 2H).

Step 3:4-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2H-benzo[b][1,4]oxazin-3(4H)-one

To a solution ofN-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-2-(2-iodophenoxy)acetamide(100 mg, 0.215 mmol) in dioxane (5 mL) was added Cs₂CO₃ (139 mg, 0.429mmol), BINAP (13.3 mg, 0.0215 mmol) and Pd₂(dba)₃ (8.0 mg, 0.0215 mmol).The reaction mixture was heated at 120° C. for 16 h, cooled to roomtemperature, filtered, and concentrated. The crude produce was purifiedby preparative HPLC purification. (23.6 mg, yield 32.5%) MS (ESI⁺) e/z:339.2 [M+1]⁺. ¹H NMR (MeOD, 400 MHz), δ ppm: 7.41-7.32 (m, 1H),7.15-7.06 (m, 3H), 7.06-6.94 (m, 4H), 4.67-4.50 (m, 2H), 4.29-4.13 (m,2H), 4.07-3.94 (m, 1H), 3.77-3.64 (m, 2H), 2.95-2.83 (m, 3H), 2.83-2.76(m, 1H), 2.73-2.62 (m, 2H).

LC-MS Conditions

Method A (LCMS-B (0-60AB_ELSD_2MIN))

Experiments performed on an Agilent 1200 HPLC (with a PDA detector and aELSD detector) with Agilent 6100 MSD mass spectrometer using ESI asionization source using an Xtimate™-C18 30*2.1 mm column and a 0.8ml/minute flow rate. Acquire Time: 2 min, Wavelength: UV220, Oven Temp.:50° C. The solvent system was a gradient starting with 100% watercontaining 0.038% TFA (solvent A) and 0% acetonitrile containing 0.02%TFA (solvent B), followed by a gradient up to 40% solvent A and 60%solvent B over the next 0.9 minutes. This was maintained for 0.6 minutesbefore returning to 100% solvent A over the next 0.5 minute. Total runtime was 2 min.

Method B (LCMS-C(10-80_AB))

Experiments performed on an SHIMADZU 20A HPLC (with a PDA detector) withSHIMADZU 2010EV MSD mass spectrometer using ESI as ionization sourceusing an Xtimate™-C18 30*2.1 mm column and a 1.2 ml/minute flow rate.The solvent system was a gradient starting with 90% water containing0.038% TFA (solvent A) and 10% acetonitrile containing 0.02% TFA(solvent B), followed by a gradient up to 20% solvent A and 80% solventB over the next 0.9 minutes. This was maintained for 0.6 minutes beforereturning to 90% solvent A and 10% solvent B over the next 0.5 minute.Total run time was 2 min.

Method C (LCMS-E(5-95AB_220&254 nm))

Experiments performed on an SHIMADZU 20A HPLC (with a PDA detector) withSHIMADZU 2010EV MSD mass spectrometer using ESI as ionization sourceusing an Merk RP-18e 2*25 mm column and a 1.5 ml/minute flow rate. Thesolvent system was a gradient starting with 95% water containing 0.038%TFA (solvent A) and 5% acetonitrile containing 0.02% TFA (solvent B),followed by a gradient up to 5% solvent A and 95% solvent B over thenext 0.7 minutes. This was maintained for 0.4 minutes before returningto 95% solvent A and 5% solvent B over the next 0.4 minute. Total runtime was 1.5 min.

Method D (LCMS-A(0-30_AB))

Experiments performed on an SHIMADZU 20A HPLC (with a PDA detector) withSHIMADZU 2010EV MSD mass spectrometer using ESI as ionization sourceusing an Xtimate™-C18 30*2.1 mm column and a 1.2 ml/minute flow rate.The solvent system was a gradient starting with 100% water containing0.038% TFA (solvent A) and 0% acetonitrile containing 0.02% TFA (solventB), followed by a gradient up to 70% solvent A and 30% solvent B overthe next 0.9 minutes. This was maintained for 0.6 minutes beforereturning to 100% solvent A over the next 0.5 minute. Total run time was2 min.

General HPLC Conditions (Acidic)

-   Mobile phase A: 4 L H₂O\1.5 ml TFA; Mobile phase B: 4 L ACN\0.75 ml    TFA-   Column: HPLC-D: Innovation C18 UPLC Column 2.1×30 mm, 2.6 um    -   HPLC-E: Xtimate C18 2.1*30 mm*3 um    -   HPLC-H: Innovation C18 UPLC Column 2.1×30 mm, 2.6 um-   Column temperature: 50° C.; Wavelength: 220 nm & 254 nm & 215 nm    General HPLC Conditions (Basic)-   Mobile phase A: 4 L H₂O\2 ml NH₄OH; Mobile phase B: Acetonitrile-   Column: HPLC-B: XBridge C18 2.1*50 mm, 5 um    -   HPLC-C: Xbridge shield RP18 2.1*50 mm, 5 u-   Column temperature: 30° C.; Wavelength: 220 nm & 254 nm & 215 nm    General HPLC Conditions (Neutral)-   Mobile phase A: H₂O; Mobile phase B: Acetonitrile-   Column: HPLC-B: XBridge C18 2.1*50 mm, 5 um    -   HPLC-C: Xbridge shield RP18 2.1*50 mm, 5 um-   Column temperature: 30° C.; Wavelength: 220 nm & 254 nm & 215 nm    Method A (0-30AB_6MIN) Flow Rate: 0.8 ml/min Gradient: 0% B to 30% B    in 4.2 min, holding 30% B for 1 min, 30% B to 0% B in 0.01 min,    holding 0% B for 1.09 min and then end.    Method B (0-60AB_6MIN)-   Flow Rate: 0.8 ml/min-   Gradient: 0% B to 60% B in 4.2 min, holding 60% B for 1 min, 60% B    to 0% B in 0.01 min, holding 0% B for 1.09 min and then end.    Method C (10-80AB_6MIN)-   Flow Rate: 0.8 ml/min-   Gradient: 10% B to 80% B in 4.2 min, holding 80% B for 1 min, 80% B    to 10% B in 0.01 min, holding 10% B for 1.09 min and then end.    Chiral HPLC Conditions:    Method A (OJ-H):    Column: Chiralcel OJ-H 250×4.6 mm I.D., 5 um    Mobile phase: A/B=90/10, A:Hexane with 0.1% DEA, B: Ethanol    Flow rate: 0.5 mL/min    Wavelength: 220 nm    Method B (OD-H):    Column: Chiralcel OD-H 250×4.6 mm I.D., 5 um    Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B: Ethanol    Flow rate: 0.5 mL/min    Wavelength: 220 nm    Method C (AD-H):    Column: Chiralpak AD-H 250×4.6 mm I.D., 5 um    Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B: Ethanol    Flow rate: 0.5 mL/min    Wavelength: 220 nm    Method D (AS-H):    Column: Chiralpak OJ-H 250×4.6 mm I.D., 5 um    Mobile phase: A/B=90/10, A: Hexane with 0.1% DEA, B: Ethanol    Flow rate: 0.5 mL/min    Wavelength: 220 nm    Biological Assays    PRMT5 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine, KCl,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG), andTris(2-carboxyethyl)phosphine hydrochloride solution (TCEP) werepurchased from Sigma-Aldrich at the highest level of purity possible.³H-SAM was purchase from American Radiolabeled Chemicals with a specificactivity of 80 Ci/mmol. 384-well streptavidin Flashplates were purchasedfrom PerkinElmer.

Substrates.

Peptide representative of human histone H4 residues 1-15 was synthesizedwith a C-terminal linker-affinity tag motif and a C-terminal amide capby 21^(st) Century Biochemicals. The peptide was high high-performanceliquid chromatography (HPLC) purified to greater than 95% purity andconfirmed by liquid chromatography mass spectrometry (LC-MS). Thesequence was Ac-SGRGKGGKGLGKGGA[K-Biot]-amide (SEQ ID NO.:3).

Molecular Biology:

Full-length human PRMT5 (NM_006109.3) transcript variant 1 clone wasamplified from a fetal brain cDNA library, incorporating flanking 5′sequence encoding a FLAG tag (MDYKDDDDK) (SEQ ID NO.:4) fused directlyto Ala 2 of PRMT5. Full-length human MEP50 (NM_024102) clone wasamplified from a human testis cDNA library incorporating a 5′ sequenceencoding a 6-histidine tag (MHHHHHH) (SEQ ID NO.:5) fused directly toArg 2 of MEP50. The amplified genes were sublconed into pENTR/D/TEV(Life Technologies) and subsequently transferred by Gateway™ attL×attRrecombination to pDEST8 baculvirus expression vector (LifeTechnologies).

Protein Expression.

Recombinant baculovirus and Baculovirus-Infected Insect Cells (BIIC)were generated according to Bac-to-Bac kit instructions (LifeTechnologies) and Wasilko, 2006, respectively. Protein over-expressionwas accomplished by infecting exponentially growing Spodopterafrugiperda (SF9) cell culture at 1.2×10⁶ cell/ml with a 5000 folddilution of BIIC stock. Infections were carried out at 27° C. for 72hours, harvested by centrifugation, and stored at −80° C. forpurification.

Protein Purification.

Expressed full-length human Flag-PRMT5/6His-MeP50 protein complex waspurified from cell paste by NiNTA agarose affinity chromatography aftera five hour equilibration of the resin with buffer containing 50 mMTris-HCL, pH 8.0, 25 mM NaCl, and 1 mM TCEP at 4° C., to minimize theadsorption of tubulin impurity by the resin. Flag-PRMT5/6His-MeP50 waseluted with 300 mM Imidazole in the same buffer. The purity of recoveredprotein was 87%. Reference: Wasilko, D. J. and S. E. Lee: “TIPS:titerless infected-cells preservation and scale-up” Bioprocess J., 5(2006), pp. 29-32.

Predicted Translations:

Flag-PRMT5 (SEQ ID NO.: 6) MDYKDDDDKA AMAVGGAGGS RVSSGRDLNC VPEIADTLGAVAKQGFDFLC MPVFHPRFKR EFIQEPAKNR PGPQTRSDLLLSGRDWNTLI VGKLSPWIRP DSKVEKIRRN SEAAMLQELNFGAYLGLPAF LLPLNQEDNT NLARVLTNHI HTGHHSSMFWMRVPLVAPED LRDDIIENAP TTHTEEYSGE EKTWMWWHNFRTLCDYSKRI AVALEIGADL PSNHVIDRWL GEPIKAAILPTSIFLTNKKG FPVLSKMHQR LIFRLLKLEV QFIITGTNHHSEKEFCSYLQ YLEYLSQNRP PPNAYELFAK GYEDYLQSPLQPLMDNLESQ TYEVFEKDPI KYSQYQQAIY KCLLDRVPEEEKDTNVQVLM VLGAGRGPLV NASLRAAKQA DRRIKLYAVEKNPNAVVTLE NWQFEEWGSQ VTVVSSDMRE WVAPEKADIIVSELLGSFAD NELSPECLDG AQHFLKDDGV SIPGEYTSFLAPISSSKLYN EVRACREKDR DPEAQFEMPY VVRLHNFHQLSAPQPCFTFS HPNRDPMIDN NRYCTLEFPV EVNTVLHGFAGYFETVLYQD ITLSIRPETH SPGMFSWFPI LFPIKQPITVREGQTICVRF WRCSNSKKVW YEWAVTAPVC SAIHNPTGRS YTIG L 6His-MEP50(SEQ ID NO.: 7) MHHHHHHRKE TPPPLVPPAA REWNLPPNAP ACMERQLEAARYRSDGALLL GASSLSGRCW AGSLWLFKDP CAAPNEGFCSAGVQTEAGVA DLTWVGERGI LVASDSGAVE LWELDENETLIVSKFCKYEH DDIVSTVSVL SSGTQAVSGS KDICIKVWDLAQQVVLSSYR AHAAQVTCVA ASPHKDSVFL SCSEDNRILLWDTRCPKPAS QIGCSAPGYL PTSLAWHPQQ SEVFVFGDENGTVSLVDTKS TSCVLSSAVH SQCVTGLVFS PHSVPFLASLSEDCSLAVLD SSLSELFRSQ AHRDFVRDAT WSPLNHSLLTTVGWDHQVVH HVVPTEPLPA PGPASVTE

General Procedure for PRMT5/MEP50 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween20, prepared on the dayof use. Compounds in 100% DMSO (1 ul) were spotted into a polypropylene384-well V-bottom plates (Greiner) using a Platemate Plus outfitted witha 384-channel head (Thermo Scientific). DMSO (1 ul) was added to Columns11, 12, 23, 24, rows A-H for the maximum signal control and 1 ul of SAH,a known product and inhibitor of PRMT5/MEP50, was added to columns 11,12, 23, 24, rows I-P for the minimum signal control. A cocktail (40 ul)containing the PRMT5/MEP50 enzyme and the peptide was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate withPRMT5/MEP50 for 30 min at 25 degrees Celsius, then a cocktail (10 ul)containing ³H-SAM was added to initiate the reaction (final volume=51ul). The final concentrations of the components were as follows:PRMT5/MEP50 was 4 nM, ³H-SAM was 75 nM, peptide was 40 nM, SAH in theminimum signal control wells was 100 uM, and the DMSO concentration was1%. The assays were stopped by the addition of non-radioactive SAM (10ul) to a final concentration of 600 uM, which dilutes the ³H-SAM to alevel where its incorporation into the peptide substrate is no longerdetectable. 50 ul of the reaction in the 384-well polypropylene platewas then transferred to a 384-well Flashplate and the biotinylatedpeptides were allowed to bind to the streptavidin surface for at least 1hour before being washed three times with 0.1% Tween20 in a BiotekELx405 plate washer. The plates were then read in a PerkinElmer TopCountplate reader to measure the quantity of ³H-labeled peptide bound to theFlashplate surface, measured as disintegrations per minute (dpm) oralternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\%\mspace{14mu}{inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu}{Coefficient}}} }}$Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.Z-138 Methylation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum, and D-PBSwere purchased from Life Technologies, Grand Island, N.Y., USA. Odysseyblocking buffer, 800CW goat anti-rabbit IgG (H+L) antibody, and LicorOdyssey infrared scanner were purchased from Licor Biosciences, Lincoln,Nebr., USA. Symmetric di-methyl arginine antibody was purchased from EMDMillipore, Billerica, Mass., USA. 16% Paraformaldehyde was purchasedfrom Electron Microscopy Sciences, Hatfield, Pa., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum and 100units/mL penicillin-streptomycin) and cultured at 37° C. under 5% CO₂.

Cell Treatment, in Cell Western (ICW) for Detection of SymmetricDi-Methyl Arginine and DNA Content.

Z-138 cells were seeded in assay medium at a concentration of 50,000cells per mL to a 384-well cell culture plate with 50 μL per well.Compound (100 nL) from 384 well source plates was added directly to 384well cell plate. Plates were incubated at 37° C., 5% CO₂ for 96 hours.After four days of incubation, 40 μL of cells from incubated plates wereadded to poly-D-lysine coated 384 well culture plates (BD Biosciences356697). Plates were incubated at room temperature for 30 minutes thenincubated at 37° C., 5% CO₂ for 5 hours. After the incubation, 40 μL perwell of 8% paraformaldehyde in PBS (16% paraformaldahyde was diluted to8% in PBS) was added to each plate and incubated for 30 minutes. Plateswere transferred to a Biotek 405 plate washer and washed 5 times with100 μL per well of wash buffer (1×PBS with 0.1% Triton X-100 (v/v)).Next 30 μL per well of Odyssey blocking buffer were added to each plateand incubated 1 hour at room temperature. Blocking buffer was removedand 20 μL per well of primary antibody was added (symmetric di-methylarginine diluted 1:100 in Odyssey buffer with 0.1% Tween 20 (v/v)) andplates were incubated overnight (16 hours) at 4° C. Plates were washed 5times with 100 μL per well of wash buffer. Next 20 μL per well ofsecondary antibody was added (1:200 800CW goat anti-rabbit IgG (H+L)antibody, 1:1000 DRAQ5 (Biostatus limited) in Odyssey buffer with 0.1%Tween 20 (v/v)) and incubated for 1 hour at room temperature. The plateswere washed 5 times with 100 μL per well wash buffer then 1 time with100 μL per well of water. Plates were allowed to dry at room temperaturethen imaged on the Licor Odyssey machine which measures integratedintensity at 700 nm and 800 nm wavelengths. Both 700 and 800 channelswere scanned.

Calculations:

First, the ratio for each well was determined by:

$( \frac{{symmetric}\mspace{14mu}{di}\text{-}{methyl}\mspace{14mu}{Arginine}\mspace{14mu} 800\mspace{14mu}{nm}\mspace{14mu}{value}}{{DRAQ}\; 5\mspace{14mu} 700\mspace{14mu}{nm}\mspace{14mu}{value}} )$

Each plate included fourteen control wells of DMSO only treatment(minimum inhibition) as well as fourteen control wells for maximuminhibition treated with 3 μM of a reference compound (Background wells).The average of the ratio values for each control type was calculated andused to determine the percent inhibition for each test well in theplate. Reference compound was serially diluted three-fold in DMSO for atotal of nine test concentrations, beginning at 3 μM. Percent inhibitionwas determined and IC₅₀ curves were generated using triplicate wells perconcentration of compound.

${{Percent}\mspace{14mu}{Inhibition}} = {100 - ( {( \frac{( {{Individual}\mspace{14mu}{Test}\mspace{14mu}{Sample}\mspace{14mu}{Ratio}} ) - ( {{Background}\mspace{14mu}{Avg}\mspace{14mu}{Ratio}} )}{( {{Minimum}\mspace{14mu}{Inhibition}\mspace{14mu}{Ratio}} ) - ( {{Background}\mspace{14mu}{Average}\mspace{14mu}{Ratio}} )} )*100} )}$Z-138 Proliferation Assay

Z-138 suspension cells were purchased from ATCC (American Type CultureCollection, Manassas, Va.). RPMI/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum werepurchased from Life Technologies, Grand Island, N.Y., USA. V-bottompolypropylene 384-well plates were purchased from Greiner Bio-One,Monroe, N.C., USA. Cell culture 384-well white opaque plates werepurchased from Perkin Elmer, Waltham, Mass., USA. Cell-Titer Glo® waspurchased from Promega Corporation, Madison, Wis., USA. SpectraMax M5plate reader was purchased from Molecular Devices LLC, Sunnyvale,Calif., USA.

Z-138 suspension cells were maintained in growth medium (RPMI 1640supplemented with 10% v/v heat inactivated fetal bovine serum andcultured at 37° C. under 5% CO₂. Under assay conditions, cells wereincubated in assay medium (RPMI 1640 supplemented with 10% v/v heatinactivated fetal bovine serum and 100 units/mL penicillin-streptomycin)at 37° C. under 5% CO₂.

For the assessment of the effect of compounds on the proliferation ofthe Z-138 cell line, exponentially growing cells were plated in 384-wellwhite opaque plates at a density of 10,000 cells/ml in a final volume of50 μl of assay medium. A compound source plate was prepared byperforming triplicate nine-point 3-fold serial dilutions in DMSO,beginning at 10 mM (final top concentration of compound in the assay was20 M and the DMSO was 0.2%). A 100 nL aliquot from the compound stockplate was added to its respective well in the cell plate. The 100%inhibition control consisted of cells treated with 200 nM finalconcentration of staurosporine and the 0% inhibition control consistedof DMSO treated cells. After addition of compounds, assay plates wereincubated for 5 days at 37° C., 5% CO₂, relative humidity>90%.

Cell viability was measured by quantitation of ATP present in the cellcultures, adding 35 μl of Cell Titer Glo® reagent to the cell plates.Luminescence was read in the SpectraMax M5 microplate reader. Theconcentration of compound inhibiting cell viability by 50% wasdetermined using a 4-parametric fit of the normalized dose responsecurves.

Results for certain compounds described herein are shown in Table 2.

TABLE 2 Biological Assay Results Biochemical Proliferation Cmpd No IC₅₀ICW EC₅₀ EC₅₀ 1 B B ** 2 D — — 3 D — — 4 A B C 5 A C — 6 A B ** 7 A B D8 A B C 9 B B D 10 — E — 11 — E — 12 A A C 13 — E — 14 — E — 15 B B **16 C — — 17 C — — 18 C — — 19 B B ** 20 B C — 21 C — — 22 C — — 23 C — —24 C — — 25 C — — 26 C — — 27 E — — 28 C — — 29 B B D 30 C — — 31 B B **32 * — — 33 B C ** 34 C — — For Table 2, “A” indicates an IC₅₀ or EC₅₀<0.100 μM, “B” indicates an IC₅₀ or EC₅₀ of 0.101-1.000 μM, “C”indicates an IC₅₀ or EC₅₀ of 1.001-10.000 μM, “D” indicates an IC₅₀ orEC₅₀ of 10.001-50 μM, and “E” indicates an IC₅₀ or EC₅₀ >50 μM. “—”indicates no data. “*” indicates an IC₅₀ or EC₅₀ >10 μM. “**” indicatesan IC₅₀ or EC₅₀ >20 μM.

Other Embodiments

The foregoing has been a description of certain non-limiting embodimentsof the invention. Those of ordinary skill in the art will appreciatethat various changes and modifications to this description may be madewithout departing from the spirit or scope of the present invention, asdefined in the following claims.

What is claimed is:
 1. A method of treating a PRMT5-mediated disorder,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond; Ring A is an optionally substituted6-membered monocyclic heterocyclyl having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur, or an optionally substituted6-membered monocyclic heteroaryl having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur; R¹ is hydrogen, R^(z), or—C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆ alkyl; Y is O;R⁵, R⁶, R⁷, and R⁸ are independently hydrogen, halo, or optionallysubstituted aliphatic; each R^(x) is independently selected from thegroup consisting of halo, —CN, optionally substituted aliphatic, —OR′,and —N(R″)₂; R′ is hydrogen or optionally substituted aliphatic; each R″is independently hydrogen or optionally substituted aliphatic, or two R″are taken together with their intervening atoms to form a heterocyclicring; and n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits;wherein, and unless otherwise specified, heterocyclyl or heterocyclicrefers to a radical of a 3-10 membered non-aromatic ring system havingring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom isindependently selected from nitrogen, oxygen, and sulfur; carbocyclyl orcarbocyclic refers to a radical of a non-aromatic cyclic hydrocarbongroup having from 3 to 10 ring carbon atoms and zero heteroatoms in thenon-aromatic ring system; aryl refers to a radical of a monocyclic orpolycyclic aromatic ring system having 6-14 ring carbon atoms and zeroheteroatoms provided in the aromatic ring system; and heteroaryl refersto a radical of a 5-10 membered monocyclic or bicyclic aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen and sulfur; each instance of aliphatic isindependently an alkyl, alkenyl, alkynyl, cycloalkyl, or cycloalkenylgroup; each instance of optionally substituted is independentlyunsubstituted or substituted, and each instance of substituted is, whenon a carbon atom, halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa),N(R^(bb))₂, —SII, —SR^(aa), —C(═O)R^(aa), —CO₂H, —CHO, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂,—C(═O)NR^(bb)SO₂R^(aa), —NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa),C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 membered heterocyclyl,C₆aryl, or 5-6 membered heteroaryl, wherein each alkyl, alkenyl,alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, or 2 R^(dd) groups, or two geminalhydrogens on the carbon atom are replaced with the group ═O; eachinstance of substituted is, when on a nitrogen, independently —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —SO₂N(R^(cc))₂, —SO₂R^(cc), C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic3-6 membered heterocyclyl, C₆aryl, or 5-6 membered heteroaryl, or twoR^(cc) groups attached to a nitrogen atom are joined to form amonocyclic 3-6 membered heterocyclyl or 5-6 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, or 2 R^(dd)groups; each instance of R^(aa) is, independently, C₁₋₁₀ alkyl, C₁₋₁₀perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆ carbocyclyl,monocyclic 3-6 membered heterocyclyl, C₆aryl, or 5-6 memberedheteroaryl, or two R^(aa) groups are joined to form a monocyclic 3-6membered heterocyclyl or 5-6 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, or 2 R^(dd) groups; eachinstance of R^(bb) is, independently, hydrogen, —OH, —OR^(aa),—N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —SO₂N(R^(cc))₂, —SO₂R^(cc), C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl,C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic3-6 membered heterocyclyl, C₆aryl, or 5-6 membered heteroaryl, or twoR^(bb)groups are joined to form a monocyclic 3-6 membered heterocyclylor 5-6 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, or 2 R^(dd) groups; each instance of R^(cc) is,independently, hydrogen, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 memberedheterocyclyl, C₆aryl, or 5-6 membered heteroaryl, or two R^(cc) groupsare joined to form a monocyclic 3-6 membered heterocyclyl or 5-6membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, or 2 R^(dd) groups; each instance of R^(dd) is,independently, halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee),—N(R^(ff))₂, —SH, —SR^(cc), —C(═O)R^(ee), —CO₂H, —CO₂R^(cc),—OC(═O)R^(ee), —OCO₂R^(ee), —C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂,—NR^(ff)C(═O)R^(cc), —NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂,—NR^(ff)SO₂R^(ee), —SO₂N(R^(ff))₂, —SO₂R^(ee), C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, monocyclic C₃₋₆ carbocyclyl,monocyclic 3-6 membered heterocyclyl, C₆aryl, or 5-6 memberedheteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl is independently substituted with 0,1, or 2 R^(gg) groups; each instance of R^(ee) is, independently, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, wherein eachalkyl, alkenyl, and alkynyl, and alkynyl is independently substitutedwith 0, 1, 2, 3, 4, or 5 R^(gg) groups; each instance of R^(ff) is,independently, hydrogen, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, orC₂₋₆ alkynyl, wherein each alkyl, alkenyl, and alkynyl, is independentlysubstituted with 0, 1, or 2 R^(gg) groups; and each instance of R^(gg)is, independently, halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OC₁₋₆alkyl, —N(C₁₋₆ alkyl)₂, —SH, —SC₁₋₆ alkyl, —C(═O)(C₁₋₆ alkyl), —CO₂H,—CO₂(C₁₋₆ alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂,—C(═O)N(C₁₋₆ alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl),—N(C₁₋₆ alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆alkyl)₂, —NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl; wherein: thePRMT5-mediated disorder is cancer, a metabolic disorder, or a blooddisorder; the metabolic disorder is diabetes or obesity; and the blooddisorder is sickle cell anemia or β-thalessemia.
 2. A method of treatinga PRMT5-mediated disorder, comprising administering to a subject in needthereof a therapeutically effective amount of a compound of Formula(II):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is hydrogen,R^(z), or —C(O)R^(z), wherein R^(z) is optionally substituted C₁₋₆alkyl; Y is O; each R^(x) is independently selected from the groupconsisting of halo, —CN, optionally substituted aliphatic, —OR′, and—N(R″)₂; R′ is hydrogen or optionally substituted aliphatic; each R″ isindependently hydrogen or optionally substituted aliphatic, or two R″are taken together with their intervening atoms to form a heterocyclicring; G is NR², CR³R⁴, O or S; R² is selected from the group consistingof optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted aryl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —C(O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(═NR^(B))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A),—C(═S)N(R^(B)), —S(═O)R^(A), —SO₂R^(A), and —SO₂N(R^(B))₂; R³ isselected from the group consisting of hydrogen, halo, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; each R^(A) is independently selectedfrom the group consisting of hydrogen, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl; eachR^(B) is independently selected from the group consisting of hydrogen,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl, or two R^(B) groups are takentogether with their intervening atoms to form an optionally substitutedheterocyclic ring; R⁴ is selected from the group consisting of hydrogen,halo, and optionally substituted aliphatic; each R^(y) is independentlyselected from the group consisting of halo, —CN, —NO₂, optionallysubstituted aliphatic, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; p is 1; m is 0, 1, 2, 3, or 4; and nis 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits; wherein, andunless otherwise specified, heterocyclyl or heterocyclic refers to aradical of a 3-10 membered non-aromatic ring system having ring carbonatoms and 1-4 ring heteroatoms, wherein each heteroatom is independentlyselected from nitrogen, oxygen, and sulfur; carbocyclyl or carbocyclicrefers to a radical of a non-aromatic cyclic hydrocarbon group havingfrom 3 to 10 ring carbon atoms and zero heteroatoms in the non-aromaticring system; aryl refers to a radical of a monocyclic or polycyclicaromatic ring system having 6-14 ring carbon atoms and zero heteroatomsprovided in the aromatic ring system; and heteroaryl refers to a radicalof a 5-10 membered monocyclic or bicyclic aromatic ring system havingring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ringsystem, wherein each heteroatom is independently selected from nitrogen,oxygen and sulfur; each instance of aliphatic is independently an alkyl,alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; each instance ofoptionally substituted is independently unsubstituted or substituted,and each instance of substituted is, when on a carbon atom, halogen,—CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —N(R^(bb))₂, —SH, —SR^(aa),—C(═O)R^(aa), —CO₂H, —CHO, —CO₂R^(aa), —OC(═O)R^(aa), —OCO₂R^(aa),—C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂, —NR^(bb)C(═O)R^(aa),—NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), C₁₋₁₀ alkyl, C₁₋₁₀perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆ carbocyclyl,monocyclic 3-6 membered heterocyclyl, C₆aryl, or 5-6 memberedheteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl is independently substituted with 0,1, or 2 R^(dd) groups, or two geminal hydrogens on the carbon atom arcreplaced with the group ═O; each instance of substituted is, when on anitrogen, independently —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —SO₂N(R^(cc))₂, —SO₂R^(cc),C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 membered heterocyclyl,C₆aryl, or 5-6 membered heteroaryl, or two R^(cc) groups attached to anitrogen atom arc joined to form a monocyclic 3-6 membered heterocyclylor 5-6 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, or 2 R^(dd) groups; each instance of R^(aa) is,independently, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 memberedheterocyclyl, C₆aryl, or 5-6 membered heteroaryl, or two R^(aa) groupsare joined to form a monocyclic 3-6 membered heterocyclyl or 5-6membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, or 2 R^(dd) groups; each instance of R^(bb) is,independently, hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —SO₂N(R^(cc))C₂, —SO₂R^(cc),C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 membered heterocyclyl,C₆aryl, or 5-6 membered heteroaryl, or two R^(bb) groups are joined toform a monocyclic 3-6 membered heterocyclyl or 5-6 membered heteroarylring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,aryl, and heteroaryl is independently substituted with 0, 1, or 2 R^(dd)groups; each instance of R^(cc) is, independently, hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆carbocyclyl, monocyclic 3-6 membered heterocyclyl, C₆aryl, or 5-6membered heteroaryl, or two R^(cc) groups are joined to form amonocyclic 3-6 membered heterocyclyl or 5-6 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, or 2 R^(dd)groups; each instance of R^(dd) is, independently, halogen, —CN, —NO₂,—N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —N(R^(ff))₂, —SH, —SR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —NR^(ff)SO₂R^(ee),—SO₂N(R^(ff))₂, —SO₂R^(ee), C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 memberedheterocyclyl, C₆aryl, or 5-6 membered heteroaryl, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, or 2 R^(gg) groups; each instanceof R^(cc) is, independently, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆alkenyl, or C₂₋₆ alkynyl, wherein each alkyl, alkenyl, and alkynyl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; eachinstance of R^(ff) is, independently, hydrogen, C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, wherein each alkyl,alkenyl, and alkynyl is independently substituted with 0, 1, or 2 R^(gg)groups; and each instance of R^(gg) is, independently, halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —N(C₁₋₆ alkyl)₂, —SH, —SC₁₋₆alkyl, —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆ alkyl), —OC(═O)(C₁₋₆ alkyl),—OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆ alkyl)₂, —OC(═O)NH(C₁₋₆alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆ alkyl)C(═O)(C₁₋₆ alkyl),—NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂, —NHC(═O)NH(C₁₋₆ alkyl),—NHC(═O)NH₂, —NHSO₂(C₁₋₆ alkyl), —SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl),—SO₂NH₂, —SO₂C₁₋₆ alkyl, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, orC₂₋₆ alkynyl; wherein: the PRMT5-mediated disorder is cancer, ametabolic disorder, or a blood disorder; the metabolic disorder isdiabetes or obesity; and the blood disorder is sickle cell anemia orβ-thalessemia.
 3. The method of claim 2, wherein the compound is ofFormula (III):

or a pharmaceutically acceptable salt thereof.
 4. The method of claim 2,wherein the compound is of Formula (IV):

or a pharmaceutically acceptable salt thereof.
 5. A method of treating aPRMT5-mediated disorder, comprising administering to a subject in needthereof a therapeutically effective amount of a compound of Formula (V):

or a pharmaceutically acceptable salt thereof; wherein: each R^(x) isindependently selected from the group consisting of halo, —CN,optionally substituted aliphatic, —OR′, and —N(R″); R′ is hydrogen oroptionally substituted aliphatic; each R″ is independently hydrogen oroptionally substituted aliphatic, or two R″ are taken together withtheir intervening atoms to form a heterocyclic ring; each R^(y) isindependently selected from the group consisting of halo, —CN, —NO₂,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted aryl, optionally substituted heterocyclyl,optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A),—C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A); —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), and —SO₂N(R^(B))₂; each R^(A) is independently selectedfrom the group consisting of hydrogen, optionally substituted aliphatic,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl; eachR^(B) is independently selected from the group consisting of hydrogen,optionally substituted aliphatic, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl, or two R^(B) groups are takentogether with their intervening atoms to form an optionally substitutedheterocyclic ring; m is 0, 1, 2, 3, or 4; and n is 0, 1, 2, 3, 4, 5, 6,7, 8, 9, or 10, as valency permits; wherein, and unless otherwisespecified, heterocyclyl or heterocyclic refers to a radical of a 3-10membered non-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur; carbocyclyl or carbocyclic refers to aradical of a non-aromatic cyclic hydrocarbon group having from 3 to 10ring carbon atoms and zero heteroatoms in the non-aromatic ring system;aryl refers to a radical of a monocyclic or polycyclic aromatic ringsystem having 6-14 ring carbon atoms and zero heteroatoms provided inthe aromatic ring system; and heteroaryl refers to a radical of a 5-10membered monocyclic or bicyclic aromatic ring system having ring carbonatoms and 1-4 ring heteroatoms provided in the aromatic ring system,wherein each heteroatom is independently selected from nitrogen, oxygenand sulfur; each instance of aliphatic is independently an alkyl,alkenyl, alkynyl, cycloalkyl, or cycloalkenyl group; each instance ofoptionally substituted is independently unsubstituted or substituted,and each instance of substituted is, when on a carbon atom, halogen,—CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —N(R^(bb))₂, —SH, —SR^(aa),—C(═O)R^(aa), CO₂H, —CHO, —CO₂R^(aa), —OC(═O)R^(aa), —OCO₂R^(aa),—C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂, —NR^(bb)C(═O)R^(aa),—NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), C₁₋₁₀ alkyl, C₁₋₁₀perhaloalkyl, C₁₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆ carbocyclyl,monocyclic 3-6 membered heterocyclyl, C₆aryl, or 5-6 memberedheteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl is independently substituted with 0,1, or 2 R^(dd) groups, or two geminal hydrogens on the carbon atom arereplaced with the group ═O; each instance of substituted is, when on anitrogen, independently —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —SO₂N(R^(cc))₂, —SO₂R^(cc),C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 membered heterocyclyl,C₆aryl, or 5-6 membered heteroaryl, or two R^(cc) groups attached to anitrogen atom are joined to form a monocyclic 3-6 membered heterocyclylor 5-6 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, or 2 R^(dd) groups; each instance of R^(aa) is,independently, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 memberedheterocyclyl, C₆aryl, or 5-6 membered heteroaryl, or two R^(aa) groupsare joined to form a monocyclic 3-6 membered heterocyclyl or 5-6membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, or 2 R^(dd) groups; each instance of R^(bb) is,independently, hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —SO₂N(R^(cc))₂, —SO₂R^(cc),C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 membered heterocyclyl,C₆aryl, or 5-6 membered heteroaryl, or two R^(bb) groups are joined toform a monocyclic 3-6 membered heterocyclyl or 5-6 membered heteroarylring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,aryl, and heteroaryl is independently substituted with 0, 1, or 2 R^(dd)groups; each instance of R^(cc) is, independently, hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, monocyclic C₃₋₆carbocyclyl, monocyclic 3-6 membered heterocyclyl, C₆aryl, or 5-6membered heteroaryl, or two R^(cc) groups are joined to form amonocyclic 3-6 membered heterocyclyl or 5-6 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, or 2 R^(dd)groups; each instance of R^(dd) is, independently, halogen, —CN, —NO₂,—N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —N(R^(ff))₂, —SH, —SR^(cc),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —NR^(ff)SO₂R^(ee),—SO₂N(R^(ff))₂, —SO₂R^(ee), C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, monocyclic C₃₋₆ carbocyclyl, monocyclic 3-6 memberedheterocyclyl, C₆aryl, or 5-6 membered heteroaryl, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, or 2 R^(gg) groups; each instanceof R^(cc) is, independently, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆alkenyl, or C₂₋₆ alkynyl, wherein each alkyl, alkenyl, and alkynyl, isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; eachinstance of R^(ff) is, independently, hydrogen, C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, wherein each alkyl,alkenyl, and alkynyl is independently substituted with 0, 1, or 2 R^(gg)groups; and each instance of R^(gg) is, independently, halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —N(C₁₋₆ alkyl)₂, —SH, —SC₁₋₆alkyl, —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆ alkyl), —OC(═O)(C₁₋₆ alkyl),—OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆ alkyl)₂, —OC(═O)NH(C₁₋₆alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆ alkyl)C(═O)(C₁₋₆ alkyl),—NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁ alkyl)₂, —NHC(═O)NH(C₁₋₆ alkyl),—NHC(═O)NH₂, —NHSO₂(C₁₋₆ alkyl), —SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl),—SO₂NH₂, —SO₂C₁₋₆ alkyl, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, orC₂₋₆ alkynyl; wherein: the PRMT5-mediated disorder is cancer, ametabolic disorder, or a blood disorder; the metabolic disorder isdiabetes or obesity; and the blood disorder is sickle cell anemia orβ-thalessemia.
 6. The method of claim 1, wherein R¹ is hydrogen.
 7. Themethod of claim 1, wherein n is
 0. 8. The method of claim 1, wherein nis
 1. 9. The method of claim 2, wherein m is
 0. 10. The method of claim2, wherein G is NR², O, or CR³R⁴, wherein R⁴ is hydrogen.
 11. The methodof claim 10, wherein O is NR², and R² is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted aliphatic,—C(═O)R^(A), or —SO₂R^(A).
 12. The method of claim 1, wherein thecompound is selected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 13. The method of claim1, wherein the disorder is cancer, and the cancer is breast cancer,esophageal cancer, bladder cancer, lymphoma, medulloblastoma, rectumadenocarcinoma, colon adenocarcinoma, gastric cancer, liver cancer,adenoid cystic carcinoma, lung adenocarcinoma, head and neck squamouscell carcinoma, brain cancer, hepatocellular carcinoma, renal cellcarcinoma, oligodendroglioma, ovarian clear cell carcinoma, ovarianserous cystadenocarcinoma, hematopoietic cancer, lung cancer, prostatecancer, melanoma, or pancreatic cancer.
 14. The method of claim 1,wherein the disorder is a metabolic disorder, and the metabolic disorderis diabetes or obesity.
 15. The method of claim 1, wherein the disorderis a blood disorder, and the blood disorder is sickle cell anemia orβ-thalessemia.
 16. The method of claim 1, wherein the compound isselected from the group consisting of:

and pharmaceutically acceptable salts thereof.